Claims:We Claim:
1. A pacifier (100) for collecting a biological sample from a subject, the pacifier (100) comprising:
a collection chamber (1) having a cavity (2) at a first end (1a), and a second end (1b) connectable to an aspiration chamber (5), wherein the cavity (2) is configured to receive the biological sample from the subject;
an opening (2a) provisioned in the cavity (2) and configured to route the biological sample received in the cavity (2) into the collection chamber (1);
a constriction (6) formed within the collection chamber (1) for supporting at least one strip (3) in the collection chamber (1); and
the at least one strip (3) provided in the collection chamber (1) such that, the at least one strip (3) extends from the first end (1a) towards the second end (1b), wherein the at least one strip (3) is configured to absorb the biological sample routed through the opening (2a).

2. The pacifier (100) as claimed in claim 1, wherein the cavity (2) is adapted to collect the biological sample from the subject, during use of the pacifier (100).

3. The pacifier (100) as claimed in claim 1, wherein the at least one strip (3) includes a hydrophilic capillary channel for absorbing the biological sample routed through the opening (2a).

4. The pacifier (100) as claimed in claim 1, wherein the aspiration chamber (5) is configured to induce vacuum within the collection chamber (1) upon actuation, for extracting the biological sample collected in the cavity (2).

5. The pacifier (100) as claimed in claim 1, wherein the at least one strip (3) is adapted to restrict backflow of the biological sample.

6. The pacifier (100) as claimed in claim 1, wherein the constriction (6) in the collection chamber (1) is formed by contraction of at least one portion of a wall (4) of the collection chamber (1).

7. The pacifier (100) as claimed in claim 6, wherein the at least one portion of the wall (4) is adapted to support the at least one strip (3) at the vicinity of the opening (2a).

8. The pacifier (100) as claimed in claim 1, wherein the aspiration chamber (5) is connected to the collection chamber (1) through a locking mechanism (8).

9. The pacifier (100) as claimed in claim 1, wherein the collection chamber (1) defines at least one vent hole (9) for discharging air from the collection chamber (1) during use of the pacifier (100).

10. The pacifier (100) as claimed in claim 1, wherein the biological sample is a saliva sample.

11. The pacifier (100) as claimed in claim 1, wherein the at least one strip (3) is a lateral flow assay.

12. A method of detecting sepsis in a saliva sample, comprising:
collecting the saliva sample from a subject into a pacifier (100) comprising at least one strip (3); and
absorbing the saliva sample by the at least one strip (3) to determine the presence or absence of a target analyte to detect the sepsis.

13. The method as claimed in claim 12, wherein the sepsis is neonatal sepsis, and the target analyte is a biomarker (10) selected from a group comprising Alpha-2-macroglobulin, Alpha-6 Type IV collagen, Alpha-amylase, Alpha-fetoprotein, Amiloride-binding protein, Ankyrin 1, erythrocyte splice form 1, Ankyrin repeat-containing protein ASB-2, Anti-phospholipid immunoglobulin light chain, Anti-pneumococcal antibody 3H1 light chain, ATP-binding cassette transporter ABC3, Beta-galactoside alpha-2, 3-sialyltransferase, Brain and muscle Ah receptor nuclear translocator-like protein, Brain-muscle-ARNT-like transcription factor 2a, Ca21/calmodulin-dependent protein kinase beta-3x, Calgranulin A, Calgranulin B (MRP-14 protein), Cathepsin L, Centrosomal Nek2-associated protein, cGMP phosphodiesterase alpha-subunit, Chaperonin GroEL precursor, Chondromodulin I, CHRNB2, Collagen alpha 1(V) chain precursor, Collagen alpha 1(VI) chain precursor, Collagen alpha 2(XI) chain precursor, Crk-associated substrate p130Cas, Cystatin B, Cystatin C, Cystatin D, Cystatin S precursor, Cystatin SA precursor, Cystatin SN precursor, Cytochrome P450 2C18, Cytokeratin 4, Cytokeratin 2P, Cytokeratin 9, DC16, Defensin alpha-1 precursor, Defensin alpha-3 precursor (neutrophil-specific), Defensin HNP-3 (chain A), Diacylglycerol kinase delta2, Disintegrin-like metalloproteinase (splice form 1), DMBT1 protein, DNMT3A protein, Dopamine receptor D1, DRIM protein, Dynamin 3, Dynein heavy chain, ER81 protein, Esophagin, 5’-3’ exoribonuclease 2 (isoform 2), FEX2, Fibrillin-2, G protein-coupled receptor kinase, Gene c-abl protein, General transcription factor iib c-terminal core domain (chain A), GTP-binding protein ARD 1, Helicase-like protein, Hepatocellular carcinoma associated protein TB6, Histatin 1 precursor, Histatin 3 precursor, Hyaluronan binding protein, ICE-LAP6, Ig alpha-1 chain C region, Immunoglobulin (chain A), Integrin alpha-4 chain precursor, Intercellular adhesion molecule 4, Landsteiner-Wiener blood group protein, Interleukin-8 receptor type B, Iodide peroxidase, Kallikrein 1, Kappa B and VDJ recombination signal sequences binding protein, Kynureninase, Lacrimal lipocalin precursor, Lipocortin I, Liprin (alpha 1), Lysozyme C precursor, Malic enzyme (chain A), MHC class II antigen, MTG16, Mucin 7 precursor, Myosin alpha heavy chain (cardiac muscle), Na, K-ATPase gamma subunit, Neutrophil gelatinase (chain A), Neutrophil peptide 3 precursor, Phospholipase C-beta-3, Salivary acidic proline-rich phosphoprotein, Salivary peroxidase, Salivary proline-rich glycoprotein precursor, Salivary proline-rich phosphoprotein, SCA2 protein, Prolactin-inducible protein, Serum albumin precursor, Sodium/phosphate cotransporter 1, Somatostatin and angiotensin-like peptide receptor, Statherin precursor, Testicular protein kinase 1, Thioredoxin, Tissue kallikrein precursor, Protein S100-A8, HBA2, Protein S100-A9, Hb subunit Beta, Hb subunit delta, fructose biphosphate aldolase A, isoform 1 of serum albumin, actin-cytoplasmic 1, annexin A1, glutathione S-transferase P, Plastin-2, serotransferrin, apolipoprotein a-1, profilin-1, CRP, Serum amyloid A, procalcitonin, lipopolysaccharide binding protein, hepcidin, IL-1 beta, IL-6, IL-8, IL-10, TNF-alpha, CD64, CD11b, sCD14, E-selectin/CD62, sTREM-1, sICAM-1, Soluble urokinase-type plasminogen activator receptor (SuPAR), RANTES, alpha-1 antitrypsin, Presepsin, and combinations thereof and combinations thereof.
14. The method as claimed in claim 12, and wherein the pacifier (100) is as defined in claims 1-11, and wherein the method comprises qualitative determination, quantitative determination or a combination thereof of the target analyte.

15. The method as claimed in claim 12, wherein the quantitative determination comprises determining the concentration of the target analyte in the saliva sample by colorimetric analysis.

16. A kit (101) for detection of sepsis in a saliva sample, comprising the pacifier (100) as defined in claims 1-11 and an instruction manual comprising steps for detecting the sepsis.
, Description:
TECHNICAL FIELD
Present disclosure generally relates to a field of medical devices. Particularly, but not exclusively the present disclosure relates to a pacifier for collecting biological sample from a subject. Further, embodiments of the present disclosure discloses a method for detecting sepsis from a saliva sample of the subject and a kit for detection of the sepsis in the saliva sample.

BACKGROUND OF THE DISCLOSURE
Biological samples including but not limiting to blood samples, urine/stool samples etc., are collected for diagnosing a subject for an infection or disease. The biological samples are usually collected using invasive techniques by insertion of instruments such as but not limiting to syringes and other invasive tools which requires puncturing skin of the subject. Use of invasive techniques, may result in trauma and acute pain to the subjects, such as but not limiting to infants and elderly subjects. Further, a skilled medical practitioner and a sterile laboratory establishment becomes a mandatory requirement for performing the invasive procedures.

Owning to complexities and stringent requirements involved in employing invasive procedures for collection biological samples, collection of saliva samples for diagnosis, particularly from neonatal and pediatric subjects is gaining prominence. This is due to the fact that, collection of saliva sample may be carried out by non-invasive procedures and also saliva sample has been found to be a reliable carrier of analyte indicators.

Advancements in the field of medicine have employed use of devices or apparatuses for effectively collecting saliva samples from the subjects. One such devices or apparatuses employed for collecting saliva sample from the subject is by use of a swab. The swab includes an absorbent pad adhered to a spatula. The swab is directly inserted into mouth of the subject for collecting saliva sample.

However, this device is primitive and is best suited for scooping saliva sample from the subject’s mouth, which may cause discomfort to the subject. Thus, this device may be only usable on adult subjects and may not be suitable for neonatal and infants due to smaller size of their mouths and their inability to comply with instructions. Additionally, neonatal and infant subjects may not have continuous generation of saliva sample, and hence it becomes a difficult proposition to collect saliva sample from these subjects. Additionally, the swab or the spatula may detach during the scooping process, which may cause a choke hazard. Further, the device should also be sterile and all the parts in the device must be designed to prevent damage to oral cavities. Also, upon scooping saliva sample from the subject, the device may be exposed to bacterial contamination and thus sterility of the sample collected becomes important.

To overcome these limitations of the saliva collection devices, particularly in infants, pacifiers are employed for collecting the saliva sample. The pacifier is inserted into the infant’s mouth, wherein the saliva sample gets deposited on the surface of the pacifier, when infants starts to suck on the pacifier, which is later swabbed and collected. However, this procedure may be unable to collect sufficient saliva sample from infants. In other words, the pacifier inserted does not provide an option for assisted salivary collection for infants who are yet to develop reflex for sucking. Additionally, in some cases infants may not produce sufficient amount of saliva for salivary collection due to prematurity or any other pre-existing condition. .

The present disclosure is directed to address one or more problems as discussed above.

The information disclosed in this background section is only for enhancement of understanding of the general background of the disclosure and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.

SUMMARY OF THE DISCLOSURE
One or more drawbacks of conventional systems and process for a method for improving yield strength of a workpiece and an apparatus are overcome, and additional advantages are provided through the apparatus and a method as claimed in the present disclosure. Additional features and advantages are realized through the technicalities of the present disclosure. Other embodiments and aspects of the disclosure are described in detail herein and are considered to be a part of the claimed disclosure.

In one non-limiting embodiment of the present disclosure, a pacifier for collecting a biological sample from a subject is disclosed. The pacifier comprises a collection chamber having a cavity at a first end and a second end connectable to an aspiration chamber, wherein the cavity is configured to receive the biological sample. An opening is provisioned in the cavity and configured to route the biological sample received in the cavity into the collection chamber. A constriction is formed within the collection chamber for supporting at least one strip in the collection chamber. The at least one strip is provided in the collection chamber such that, the at least one strip extends from the first end towards the second end, wherein the at least one strip is configured to absorb the biological sample routed through the opening.

In an embodiment, the cavity is adapted to collect the biological sample from the subject, during use of the pacifier.

In an embodiment, the at least one strip includes a hydrophilic capillary channel for absorbing the biological sample collected in the cavity.

In an embodiment, the aspiration chamber is configured to induce vacuum within the collection chamber upon actuation, for extracting the biological sample collected in the cavity.

In an embodiment, the at least one strip is adapted to restrict backflow of the biological sample.

In an embodiment, the constriction in the collection chamber is formed by contraction of at least one portion of the wall of the collection chamber.

In an embodiment, the at least one portion of the wall is adapted to support the at least one strip at the vicinity of the opening.

In an embodiment, the aspiration chamber is connected to the collection chamber through a locking mechanism.

In an embodiment, the collection chamber defines at least one vent hole for discharging air from the collection chamber during use of the pacifier.

In an embodiment, the biological sample is saliva.

In an embodiment, the at least one strip is a lateral flow assay.

In an embodiment, a method of detecting sepsis in a saliva sample is disclosed. The method comprises collecting the saliva sample from a subject into a pacifier comprising at least one strip and absorbing the saliva sample by the at least one strip to determine the presence or absence of a target analyte to detect the sepsis.

In an embodiment, the sepsis is neonatal sepsis, and the target analyte is a biomarker selected from a group comprising Alpha-2-macroglobulin, Alpha-6 Type IV collagen, Alpha-amylase, Alpha-fetoprotein, Amiloride-binding protein, Ankyrin 1, erythrocyte splice form 1, Ankyrin repeat-containing protein ASB-2, Anti-phospholipid immunoglobulin light chain, Anti-pneumococcal antibody 3H1 light chain, ATP-binding cassette transporter ABC3, Beta-galactoside alpha-2, 3-sialyltransferase, Brain and muscle Ah receptor nuclear translocator-like protein, Brain-muscle-ARNT-like transcription factor 2a, Ca21/calmodulin-dependent protein kinase beta-3x, Calgranulin A, Calgranulin B (MRP-14 protein), Cathepsin L, Centrosomal Nek2-associated protein, cGMP phosphodiesterase alpha-subunit, Chaperonin GroEL precursor, Chondromodulin I, CHRNB2, Collagen alpha 1(V) chain precursor, Collagen alpha 1(VI) chain precursor, Collagen alpha 2(XI) chain precursor, Crk-associated substrate p130Cas, Cystatin B, Cystatin C, Cystatin D, Cystatin S precursor, Cystatin SA precursor, Cystatin SN precursor, Cytochrome P450 2C18, Cytokeratin 4, Cytokeratin 2P, Cytokeratin 9, DC16, Defensin alpha-1 precursor, Defensin alpha-3 precursor (neutrophil-specific), Defensin HNP-3 (chain A), Diacylglycerol kinase delta2, Disintegrin-like metalloproteinase (splice form 1), DMBT1 protein, DNMT3A protein, Dopamine receptor D1, DRIM protein, Dynamin 3, Dynein heavy chain, ER81 protein, Esophagin, 5’-3’ exoribonuclease 2 (isoform 2), FEX2, Fibrillin-2, G protein-coupled receptor kinase, Gene c-abl protein, General transcription factor iib c-terminal core domain (chain A), GTP-binding protein ARD 1, Helicase-like protein, Hepatocellular carcinoma associated protein TB6, Histatin 1 precursor, Histatin 3 precursor, Hyaluronan binding protein, ICE-LAP6, Ig alpha-1 chain C region, Immunoglobulin (chain A), Integrin alpha-4 chain precursor, Intercellular adhesion molecule 4, Landsteiner-Wiener blood group protein, Interleukin-8 receptor type B, Iodide peroxidase, Kallikrein 1, Kappa B and VDJ recombination signal sequences binding protein, Kynureninase, Lacrimal lipocalin precursor, Lipocortin I, Liprin (alpha 1), Lysozyme C precursor, Malic enzyme (chain A), MHC class II antigen, MTG16, Mucin 7 precursor, Myosin alpha heavy chain (cardiac muscle), Na, K-ATPase gamma subunit, Neutrophil gelatinase (chain A), Neutrophil peptide 3 precursor, Phospholipase C-beta-3, Salivary acidic proline-rich phosphoprotein, Salivary peroxidase, Salivary proline-rich glycoprotein precursor, Salivary proline-rich phosphoprotein, SCA2 protein, Prolactin-inducible protein, Serum albumin precursor, Sodium/phosphate cotransporter 1, Somatostatin and angiotensin-like peptide receptor, Statherin precursor, Testicular protein kinase 1, Thioredoxin, Tissue kallikrein precursor, Protein S100-A8, HBA2, Protein S100-A9, Hb subunit Beta, Hb subunit delta, fructose biphosphate aldolase A, isoform 1 of serum albumin, actin-cytoplasmic 1, annexin A1, glutathione S-transferase P, Plastin-2, serotransferrin, apolipoprotein a-1, profilin-1, CRP, Serum amyloid A, procalcitonin, lipopolysaccharide binding protein, hepcidin, IL-1 beta, IL-6, IL-8, IL-10, TNF-alpha, CD64, CD11b, sCD14, E-selectin/CD62, sTREM-1, sICAM-1, Soluble urokinase-type plasminogen activator receptor (SuPAR), RANTES, alpha-1 antitrypsin, Presepsin, and combinations thereof and combinations thereof.

In an embodiment, the method comprises qualitative determination, quantitative determination or a combination thereof of the target analyte.

In an embodiment, the quantitative determination comprises determining the concentration of the target analyte in the saliva sample by colorimetric analysis.

In another non-limiting embodiment of the present disclosure, a kit for detection of sepsis in a saliva sample is disclosed. The kit comprises the pacifier and an instruction manual comprising steps for detecting the sepsis.

It is to be understood that the aspects and embodiments of the disclosure described above may be used in any combination with each other. Several of the aspects and embodiments may be combined to form a further embodiment of the disclosure.

The foregoing summary is illustrative only and is not intended to be in any way limiting. In addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features will become apparent by reference to the drawings and the following detailed description.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS
The novel features and characteristics of the disclosure are set forth in the appended description. The disclosure itself, however, as well as a preferred mode of use, further objectives and advantages thereof, will best be understood by reference to the following detailed description of an illustrative embodiment when read in conjunction with the accompanying figures. One or more embodiments are now described, by way of example only, with reference to the accompanying figures wherein like reference numerals represent like elements and in which:

Figure 1 illustrates a sectional view of the pacifier of Figure 1, in accordance with an embodiment of the present disclosure.

Figure 2 illustrates an exploded view of a pacifier for collecting biological samples, in accordance with an embodiment of the present disclosure.

Figure 3a illustrates a side view of a collection chamber of the pacifier of Figure 1, in accordance with an embodiment of the present disclosure.

Figure 3b illustrates an enlarged view of section A of Figure 3a to illustrate configuration of a cavity in the collection chamber, in accordance with an embodiment of the present disclosure.

Figure 4 illustrates a schematic view of a kit including the pacifier of Figure 1, in accordance with an embodiment of the present disclosure.

The figures depict embodiments of the disclosure for purposes of illustration only. One skilled in the art will readily recognize from the following description that alternative embodiments of the structures and methods illustrated herein may be employed without departing from the principles of the disclosure described herein.

DETAILED DESCRIPTION OF THE DISCLOSURE

While the embodiments of the disclosure are subject to various modifications and alternative forms, specific embodiment thereof have been shown by way of example in the figures and will be described below. It should be understood, however, that it is not intended to limit the disclosure to the particular forms disclosed, but on the contrary, the disclosure is to cover all modifications, equivalents, and alternative falling within the scope of the disclosure.

It is to be noted that a person skilled in the art would be motivated from the present disclosure to arrive at a pacifier for collecting biological sample, a method of detecting sepsis and a kit thereof. The pacifier, the method of detecting sepsis and the kit may vary based on configuration of the pacifier. However, such modifications should be construed within the scope of the disclosure. Accordingly, the drawings illustrate only those specific details that are pertinent to understand the embodiments of the present disclosure, so as not to obscure the disclosure with details that will be clear to those of ordinary skill in the art having benefit of the description herein.

The terms “comprises”, “comprising”, or any other variations thereof used in the disclosure, are intended to cover a non-exclusive inclusion, such that a device, a system, an assembly that comprises a list of components does not include only those components but may include other components not expressly listed or inherent to such system, or assembly, or device. In other words, one or more elements in a system or device proceeded by “comprises… a” does not, without more constraints, preclude the existence of other elements or additional elements in the system or device.

The present disclosure provides a pacifier for collecting biological samples from a subject, including but not limiting to neonatal and premature infants. The pacifier comprises a collection chamber, having a cavity at a first end and a second end connectable to an aspiration chamber. The cavity is configured to collect the biological sample, so that the collected biological sample may be sourced for diagnosis of the subject. An opening is provided in the cavity, for routing the biological sample into the collection chamber. The aspiration chamber upon actuation is configured to induce vacuum in the collection chamber, thereby enabling drawing of biological sample into the cavity. Further, at least one strip is provided in the collection chamber wherein the at least one strip extends from the first end of the collection chamber towards the second end of the collection chamber, and is configured to absorb the biological sample received at the cavity. A constriction is formed at the first end within the collection chamber to support the at least one strip in the collection chamber, so that the at least one strip is aligned with the cavity during use of the pacifier.

The present disclosure is configured to collect biological samples from the subject, particularly from neonatal and premature infants, who are unable to secrete sufficient saliva and where prior art methods of extracting saliva is not possible. The present disclosure also enables collection of the biological sample in a sterile and hygienic manner.

The following paragraphs describe the present disclosure with reference to figures 1-4. In the figures, the same element or elements which have similar functions are indicated by the same reference signs.

Figure 1 in one exemplary embodiment of the present disclosure illustrates a sectional view of a pacifier (100), for collecting a biological sample from a subject.

The pacifier (100) comprises a collection chamber (1) configured to collect the biological sample from the subject. In an exemplary embodiment, the biological sample is saliva. The collection chamber (1) includes a cavity (2) at its first end (1a), which is configured to collect the biological sample [herein referred to as saliva] from the subject during use of the pacifier (100). This configuration ensures that the required quantity of saliva is collected in the cavity (2) during use of the pacifier (100), for evaluation. The cavity (2) includes an opening (2a) for routing the saliva from the subject into the collection chamber (1) during use of the pacifier (100).

The pacifier (100) further includes at least one strip (3) provisioned in the collection chamber (1) such that, the at least one strip (3) extends from the first end (1a) towards the second end (1b) [as shown in Figure 2]. The at least one strip (3) is configured to absorb the saliva received at the cavity (2). The at least one strip (3) is positioned adjacent to the opening (2a) so that, the saliva retained in the cavity (2) is absorbed or wicked in the at least one strip (3). The at least one strip (3) is configured with a hydrophilic capillary channel [not shown in Figures] for absorbing the saliva collected in the cavity (2).

The collection chamber (1) is defined by a wall (4) from the first end (1a) up to a second end (1b) [shown in figure 1] . The collection chamber (1) includes a narrow portion (7a) towards the first end (1a) and an enlarged portion (7b) towards the second end. A constriction (6) divides the narrow portion (7a) and the enlarged portion (7b) of the collection chamber (1). The constriction (7) is configured for supporting the at least one strip (3) in the collection chamber (1). The constriction (6) defines a depression (7c) in at least a portion of the wall (4), which is sufficient enough for axial movement of the at least one strip (3), while preventing radial movement. This configuration ensures that the position of at least one strip (3) remains adjacent to the opening (2a), thereby enable wicking or absorbing of the saliva by the at least one strip (3). Further, any other embodiment which would enable the strip (3) to be held in the vicinity of the cavity (2) to facilitate collection of saliva from the cavity (2) falls within the purview of this disclosure. In an embodiment, the narrow portion (7a) of the wall (4) is defined to be a nipple portion and the enlarged portion (7b) is defined to be an areola portion.

Further, the collection chamber (1) has the second end (1b) connectable to an aspiration chamber (5). The aspiration chamber (5) upon actuation, is configured to induce vacuum within the collection chamber (1) for extracting saliva collected in the cavity (2). This provision ensures that the saliva is collected in the cavity (2) and also enables extraction of the saliva even from subjects, who are unable to secrete sufficient saliva. In an embodiment, the aspiration chamber (5) is a hollow chamber, which induces vacuum within the collection chamber (1) due to venturi effect. The venturi effect is induced due to decrease in the cross-section of the aspiration chamber (5) upon actuation of the aspiration chamber (5). In an exemplary embodiment, the at aspiration chamber (5) is selected from at least one of an aspirator, or any other vacuum inducing device which serves the purpose.

The aspiration chamber (5) is connectable to the second end (1b) of the collection chamber (1) through a locking mechanism (8). The locking mechanism (8) forms a sealing joint between the collection chamber (1) and the aspiration chamber (5). The locking mechanism (8) also ensures ingress protection to the collection chamber (1), thereby providing sterile and hygienic conditions for collection of the saliva. In an embodiment, the locking mechanism (8) is selected from at least one of snap-fit mechanism, a linkage mechanism or any other mechanism, which serves the purpose.

In an embodiment, the collection chamber (1) is a hollow member, configured to receive the at least one strip (3).

In an embodiment, size of the cavity (2) is configured such that, sufficient quantity of the saliva for diagnosis of the subject is retained.

In an embodiment, the cavity (2) may be a cut-out or a projection provided on the pacifier (100). In an embodiment, the cross-section of the cavity (2) may be selected from at least one of a circular cross-section, rectangular cross-section or any other cross-section which serves the purpose of retaining the saliva.

In an embodiment, the opening (2a) may be provided at any position in the cavity (2). In an exemplary embodiment, the opening (2a) may be provided at an aft end portion of the cavity (2), so that the collected saliva is routed to the opening (2a) provided in the collection chamber (1).

In an embodiment, the cross-section of the opening (2a) may be a circular cross-section, a rectangular cross-section or any other cross-section which serves the purpose of routing the collected saliva into the collection chamber (1).

In an embodiment, the opening (2a) is an orifice for routing the collected saliva into the collection chamber (1). In another embodiment, the orifice [shown in figures 3a and 3b] may be a cut-out or a slot or a slit (2b) provided to the cavity (2) to route the saliva into the opening (2a) of the collection chamber (1).

In an embodiment, the at least one strip (3) may include an absorbent pad [not shown in the Figures] provided adjacent to the first end (1a) for wicking or absorbing the saliva from the opening (2a).

In an embodiment, the at least one strip (3) is configured with a plurality of layers [not shown in the Figures], laminated over each other. The plurality of layers are laminated so that the hydrophilic capillary channel is defined in the at least one strip (3). In an exemplary embodiment, the dimensions of the hydrophilic channel is in the range of about 0.5mm to about 2mm. These dimensions of the hydrophilic channel increases flow rate of the saliva entering the at least one strip (3), thereby enabling wicking or collection of the saliva. The hydrophilic capillary channel is configured to induce capillary effect in the at least one strip (3), thereby inducing lateral flow of the saliva across the entire length of the hydrophilic capillary channel. The hydrophilic capillary channel is also configured to collect a predetermined quantity of the saliva for evaluation of the subject. In an exemplary embodiment, the hydrophilic capillary channel stores the saliva in the range of about 5µL to about 20µL. This configuration of the at least one strip (3) restricts backflow of the saliva entering the collection chamber (1), thereby ensures that the saliva remains in the collection chamber (1) and also prevents contamination of the saliva due to exposure to the surroundings.

In an embodiment, the at least one strip (3) is configured with a plurality of biomarkers (10) [as shown in Figure 2] at predetermined locations along its span. This provision enables, immediate evaluation of the infection or the disease using saliva, thereby mitigating the need for complex laboratory instruments. In an embodiment, the at least one strip (3) alters its color characteristics upon detecting a particular infection or disease.

In an embodiment, the enlarged portion (7b) of the collection chamber (1) behaves as a guard of the pacifier (100), so that only the narrow portion (7a) enters the subject’s mouth. This configuration, therefore ensures that the aspiration chamber (5) acts as a holder for inserting the pacifier (100) into the subject’s mouth.

In another embodiment, at least one vent hole (9) is provided in the collection chamber (1), for discharging air during use of the pacifier (100). In an exemplary embodiment, the at least one vent hole (9) is provided at the vicinity of the second end (1b) of the collection chamber (1). The vent hole (9) also aids in creating a vacuum while operating the aspiration chamber (5).

In an embodiment, the pacifier (100) may be a single piece construction, or multi-piece construction joined together by the locking mechanism (8). In yet another embodiment, the material used for the pacifier (100) is selected from at least one of rubber, silicone or any other material which serves the purpose.

In an operational embodiment, the pacifier (100) is introduced into mouth of the subject, such that the first end (1a) of the collection chamber (1) enters the subject’s mouth while the aspiration chamber (5) remains outside the subject’s mouth. As the subject begins to suck the pacifier (100), saliva gets smeared on outer surface of the collection chamber (1). Subsequently, the saliva contacts the cavity (2) during use of the pacifier (100) and thereby ensures that the cavity (2) retains a portion of the saliva in it. The at least one strip (3) positioned adjacent to the cavity (2), wicks the saliva through the opening (2a), and collects sufficient quantity of the saliva for diagnosis of the subject.

In another embodiment, for subjects who are unable to secrete sufficient saliva, the aspiration chamber (5) is actuated and the first end (1a) is inserted into the subject’s mouth. Actuation of the aspiration chamber (5) induces vacuum in the collection chamber (1), which inherently induces vacuum at the first end (1a). The vacuum induced in the collection chamber (1), routes the saliva that contacts the first end (1a) into the collection chamber (1) through the opening (2a), thereby extracting the saliva.

The present disclosure further relates to a method of detecting sepsis in a saliva sample by employing the pacifier (100) described herein.

In an exemplary embodiment, the method of detecting sepsis comprises:
i) aspiration of saliva using the pacifier (100) as described herein;
ii) wicking of saliva directly by lateral flow assay i.e. the at least one strip (3) of the pacifier (100);
iii) colorimetric analysis for qualitative and quantitative determination of biomarker (10) of interest using a smartphone application/algorithm and corresponding calibration card.
iv) comparison of the biomarker (10) quantities obtained with smartphone application versus a predefined range to detect the sepsis.

In an embodiment, the plurality of biomarkers (10) selected from a group comprising Alpha-2-macroglobulin, Alpha-6 Type IV collagen, Alpha-amylase, Alpha-fetoprotein, Amiloride-binding protein, Ankyrin 1, erythrocyte splice form 1, Ankyrin repeat-containing protein ASB-2, Anti-phospholipid immunoglobulin light chain, Anti-pneumococcal antibody 3H1 light chain, ATP-binding cassette transporter ABC3, Beta-galactoside alpha-2, 3-sialyltransferase, Brain and muscle Ah receptor nuclear translocator-like protein, Brain-muscle-ARNT-like transcription factor 2a, Ca21/calmodulin-dependent protein kinase beta-3x, Calgranulin A, Calgranulin B (MRP-14 protein), Cathepsin L, Centrosomal Nek2-associated protein, cGMP phosphodiesterase alpha-subunit, Chaperonin GroEL precursor, Chondromodulin I, CHRNB2, Collagen alpha 1(V) chain precursor, Collagen alpha 1(VI) chain precursor, Collagen alpha 2(XI) chain precursor, Crk-associated substrate p130Cas, Cystatin B, Cystatin C, Cystatin D, Cystatin S precursor, Cystatin SA precursor, Cystatin SN precursor, Cytochrome P450 2C18, Cytokeratin 4, Cytokeratin 2P, Cytokeratin 9, DC16, Defensin alpha-1 precursor, Defensin alpha-3 precursor (neutrophil-specific), Defensin HNP-3 (chain A), Diacylglycerol kinase delta2, Disintegrin-like metalloproteinase (splice form 1), DMBT1 protein, DNMT3A protein, Dopamine receptor D1, DRIM protein, Dynamin 3, Dynein heavy chain, ER81 protein, Esophagin, 5’-3’ exoribonuclease 2 (isoform 2), FEX2, Fibrillin-2, G protein-coupled receptor kinase, Gene c-abl protein, General transcription factor iib c-terminal core domain (chain A), GTP-binding protein ARD 1, Helicase-like protein, Hepatocellular carcinoma associated protein TB6, Histatin 1 precursor, Histatin 3 precursor, Hyaluronan binding protein, ICE-LAP6, Ig alpha-1 chain C region, Immunoglobulin (chain A), Integrin alpha-4 chain precursor, Intercellular adhesion molecule 4, Landsteiner-Wiener blood group protein, Interleukin-8 receptor type B, Iodide peroxidase, Kallikrein 1, Kappa B and VDJ recombination signal sequences binding protein, Kynureninase, Lacrimal lipocalin precursor, Lipocortin I, Liprin (alpha 1), Lysozyme C precursor, Malic enzyme (chain A), MHC class II antigen, MTG16, Mucin 7 precursor, Myosin alpha heavy chain (cardiac muscle), Na, K-ATPase gamma subunit, Neutrophil gelatinase (chain A), Neutrophil peptide 3 precursor, Phospholipase C-beta-3, Salivary acidic proline-rich phosphoprotein, Salivary peroxidase, Salivary proline-rich glycoprotein precursor, Salivary proline-rich phosphoprotein, SCA2 protein, Prolactin-inducible protein, Serum albumin precursor, Sodium/phosphate cotransporter 1, Somatostatin and angiotensin-like peptide receptor, Statherin precursor, Testicular protein kinase 1, Thioredoxin, Tissue kallikrein precursor, Protein S100-A8, HBA2, Protein S100-A9, Hb subunit Beta, Hb subunit delta, fructose biphosphate aldolase A, isoform 1 of serum albumin, actin-cytoplasmic 1, annexin A1, glutathione S-transferase P, Plastin-2, serotransferrin, apolipoprotein a-1, profilin-1, CRP, Serum amyloid A, procalcitonin, lipopolysaccharide binding protein, hepcidin, IL-1 beta, IL-6, IL-8, IL-10, TNF-alpha, CD64, CD11b, sCD14, E-selectin/CD62, sTREM-1, sICAM-1, Soluble urokinase-type plasminogen activator receptor (SuPAR), RANTES, alpha-1 antitrypsin, Presepsin, and combinations thereof.

In a preferred embodiment, the biomarker is selected from a group comprising CRP, IL8, IL6, IL10, LBP, sTREM1 and combinations thereof.

In another embodiment, the method of the present disclosure further comprises understanding the effectiveness of treatment (after the patient is detected with the sepsis and therapy/treatment is undertaken). In an exemplary embodiment, understanding the effectiveness of treatment comprises:
v) calculation of a weighted score based on biomarker quantities obtained in the method described above;
vi) repeating steps i) – v) after the subject has begun treatment;
vii) comparison of weighted scores as obtained in step vi) with values as obtained in step v) to understand the effectiveness of therapy.

The present disclosure also provides a kit for detecting sepsis in a saliva sample.

In an embodiment, the kit comprises one pacifier (100) and at least one single use strip [as shown in Figure 4], along with an instruction manual for use.

In a preferred embodiment, the kit comprises one pacifier (100) and 3 single use strips.

In another embodiment, the instruction manual of the kit comprises steps to be followed for detection of sepsis in the sample.

EXAMPLE 1: Method of detecting sepsis
The method of detecting sepsis in the saliva sample by employing the pacifier (100) described herein is performed. The method involves the following steps:
i) aspiration of saliva using the pacifier (100) as described herein;
ii) wicking of saliva directly by lateral flow assay i.e. the at least one strip (3) of the pacifier (100);
iii) colorimetric analysis for qualitative and quantitative determination of biomarkers (10) of interest using a smartphone application/algorithm and corresponding calibration card.
iv) comparison of the biomarker quantities obtained with smartphone application versus a predefined range to detect the sepsis.

Procedure of Step iii): colorimetric readout using smartphone application/algorithm and corresponding calibration card
The following steps are performed to obtain colorimetric read out:
a) Lateral Flow Assay [strip (3)] is photographed by smartphone application;
b) Assay is scanned and cropped by the application to obtain two region of interests (ROIs): ROI 1 (region of interest 1) and ROI 2 (region of interest 2);
c) Shadow corrections on the extracted ROIs are performed, if any;
d) ROI 1 constitutes of a color scale printed at end of the strip;
e) ROI 2 constitutes of the nitrocellulose (NC) membrane area with test lines and control lines;
f) The 5 point color scale in ROI 1 is used to calculate a standard curve;
g) A moving window of 5 pixels is used to calculate y-coordinate vs average hue value from
ROI 2;
h) The plot in step g) gives peaks for test and control lines. The peak hue values for each test line is added to the hue value of control line - i.e. Hue (test line1+ control line) = hue 1; Hue (test line 2 + control line) = hue 2 and so on;
i) The obtained hue1, hue2, hue3 and so on are mapped on the standard curve to obtain respective colorimetric values.

To understand the effectiveness of treatment (after the patient is detected with the sepsis and therapy/treatment is undertaken), the following subsequent steps could be additionally performed:
v) calculation of a weighted score based on biomarker quantities obtained above;
vi) repeating steps i) – v) after the subject has begun treatment;
vii) comparison of weighted scores as obtained in step vi) with values as obtained in step v) to understand the effectiveness of therapy.

Procedure of Step v): Scoring Algorithm / calculation of weighted scores
About 4 biomarkers (10) are detected on the strip and weightages are assigned to each one of them. The weightages depend on median values of ranges that have been obtained from all the tests that have been done using the above described colorimetric readout procedure. For example, for data pertaining to the below Table, weights are calculated as:
W1 = [M_CP1-M_CN1] / M_CP1
W2 = [M_CP2-M_CN2] / M_CP2
W3 = [M_CP3-M_CN3] / M_CP3

The final score from the given test assay is calculated as:
(biomarker1_norm*W1+biomarker2_norm*W2+ biomarker3_norm*W3)/10

This score is used for giving a sepsis risk where: 0-3 indicates low risk of sepsis; 3-6 indicates moderate risk of sepsis; and 6-10 indicates high risk of sepsis.

EXAMPLE 2: A Kit assembly and functioning
The kit (101) of the present disclosure is developed which comprises one pacifier and 3 single use strips [as shown in Figure 3], along with an instruction manual for use. The user is advised to assemble the strip (3) along with the pacifier (100) and as shown in manual, and carry out tests after 0 hour, 24 hours and 72 hours of symptoms. The quantitative score obtained at these points could be used to guide antibiotic therapy. When in subject’s (newborn’s) mouth, the pacifier (100) simulates sucking reflex in the newborn promoting drool. When the nipple part is pressed by the baby during sucking action, saliva is drawn in. During the breathing cycle, air flows inside the nipple through the air vents on the aspirator chamber side avoiding backflow of saliva. During the relax cycle, this saliva now flows onto the lateral flow assay / strip inside the pacifier (100). After 5 mins, the pacifier (100) can be taken out, dismantled and read using the scoring algorithm as described under Example 1.

Advantages:
In an embodiment, the present disclosure provides a pacifier which provides assisted salivary collection from neonates who have not developed the reflex for sucking.

In an embodiment, the present disclosure provides a pacifier which provides an option for salivary collection from neonates who are unable to secrete sufficient amount of saliva due to prematurity or any other pre-existing condition.

In an embodiment, the present disclosure provides a pacifier which prevents backflow of the biological sample collected in the collection chamber, thereby maintaining sterility of the biological sample collected.

In an embodiment, the present disclosure provides a pacifier which enables extraction or collection of biological sample from subjects without discomforting the subject.

In an embodiment, the present disclosure provides a pacifier which readily determines sepsis or other disease by observing change of color of the at least one strip.

Equivalents
With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate to the context and/or application. The various singular/plural permutations may be expressly set forth herein for sake of clarity.

It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims (e.g., bodies of the appended claims) are generally intended as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "includes" should be interpreted as "includes but is not limited to," etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles "a" or "an" limits any particular claim containing such introduced claim recitation to inventions containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and indefinite articles such as "a" or "an" (e.g., "a" and/or "an" should typically be interpreted to mean "at least one" or "one or more"); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should typically be interpreted to mean at least the recited number (e.g., the bare recitation of "two recitations," without other modifiers, typically means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention analogous to "at least one of A, B, and C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, and C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). In those instances, where a convention analogous to "at least one of A, B, or C, etc." is used, in general such a construction is intended in the sense one having skill in the art would understand the convention (e.g., "a system having at least one of A, B, or C" would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together, etc.). It will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description, claims, or drawings, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" will be understood to include the possibilities of "A" or "B" or "A and B."

While various aspects and embodiments have been disclosed herein, other aspects and embodiments will be apparent to those skilled in the art. The various aspects and embodiments disclosed herein are for purposes of illustration and are not intended to be limiting, with the true scope and spirit being indicated by the following claims.

REFERRAL NUMERALS:

Referral numerals
Description
100 Pacifier
101 Kit
1 Collection chamber
1a First end of collection chamber
1b Second end of collection chamber
2 Cavity
2a Opening in cavity
2b Slits
3 At least one strip
4 Wall of the collection chamber
5 Aspiration chamber
6 Constriction in the collection chamber
7a Narrow portion in the collection chamber
7b Enlarged portion in the collection chamber
7c Depression in the wall
8 Locking mechanism
9 Vent holes
10 Biomarkers