FIELD OF THE INVENTION
The present invention relates to a nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR). The invention relates to a process for selecting and/or obtaining nucleic acid probes capable of detecting existing/new variable number tandem repeat regions present in the genomes of higher eucaryotes, probes obtainable in this way, and their applications. The present invention includes identification of novel VNTR sequences and use of these novel sequences as probe for cell line authentication and for identity consanguinity testing.
BACKGROUND OF THE INVENTION AND PRIOR ART
With the development of molecular biology and its application to daily life, circumstances requiring genetic identification are increasingly frequent. Among these, identification of an individual, particularly in police investigations or paternity testing, identification of a cell line, or checking the origin of a human, animal, or plant seed may be cited. However, in view of the complexity of the genomes in higher eukaryotes, a complete genome analysis for a given individual cannot be contemplated at the present time.
Direct analysis of DNA has revealed extremely polymorphic areas of the genome. Free of the constraints imposed upon variability within the expressed region of the genome, parts of the nonexpressed sequences have been discovered that have hundreds of variants. The first of these hypervariable regions (HVRs) was identified in 1980, although the structural basis for the observed variability was not known at the time. Subsequently, many other HVRs were identified, and it became clear that families of HVRs were dispersed throughout the genome.
All of the minisatellite loci detected thus far consist of tandem repeats of sequences that differ from one locus to another, but all show homology to the core sequence. The extreme degree of polymorphism observed at these loci arises from variation in the number of times the minisatellite sequence is repeated. This type of hypervariable region is also known as a VNTR (variable number of tandem repeats). The various VNTRs of a given individual are dispersed throughout the genome and are distinguished from each other, not only by the sequence of the repeated motif, but

also by the number of repetitions of their own motif Amplification of these minisatellite regions or VNTR sequences at relatively high stringencies have resulted in a PCR based methodology named as Direct Amplfication Minisatellite DNA (DAMD).
Of the methods for detecting these regions used to date, one may cite the screening of a bank using nucleotide probes with natural tandem repeats (Wong et al., Ann. Hum. Genet., 51, 269-288, 1987) or synthetic probes (Zischler et al.. Genomics, 13, 983-990, 1992). However, these methods have proved inadequate in that, thus far, they have enabled documentation of only about 10% of the estimated number of VNTRs for the human species. Development of a module is very important that could identify VNTR loci and amplifying these regions with DAMD technique and generating the signature pattern of first tier for each of the cell line/individual. Further identifying the specific band with in the signature pattern generated which can generate a unique pattern in southern (second tier pattern) when used as probe in southern can establish the identity of the individual/cell line or genome.
Extensive usage of eukaryotic cell line in manufacturing and developing biogeneric and biosimilars have agumented the need for cell line characterization where in cell line identity and the authenticity is a primary concern for the manufacutrer and as well as the regulatory authorities. Hence it became necessity to resort in using markers of this genetic identity to characterize cell lines.
DAMD is a simple and easy method to perform and the fingerprints could be used in the construction of a database for cell line control. Availability of this database to laboratories involved in cell culturing would allow easy identification of individual lines by comparison to reference profiles as well as comparison of similar lines from different sources and periodic follow-up of cells in culture.
STATEMENT OF THE INVENTION
Accordingly, the present invention relates to a nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR); a method for obtaining nucleic acid probe or its equivalent probe capable of detecting Variable

Number Tandem Repeat (VNTR), wherein said method comprises steps of: (i) identifying region of genomic DNA containing VNTR locus to get unique electrophoretic pattern; (ii) repeating step (i) on similar sample of genomic DNA to obtain consistent electrophoretic pattern; (iii) comparing the pattern obtained in step (ii) with different genomic DNA to obtain unique band; and (iv) extracting the unique band of step (iii) to obtain said probe and use of nucleic acid probe or its equivalent probe for plurality of purposes selected from a group comprising consanguinity search, screening for tumoral disease, screening for hereditary disease and identification of biological specimen/cell line or a combination thereof.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR).
In another embodiment of the present invention, said equivalent probe is selected from a group comprising natural probe, synthetic probe and semi-synthetic probe or a combination thereof and said modification comprises modification of one or more base, preferably replacing a base with inosine.
The present invention also relates to a method for obtaining nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR), wherein said method comprises steps of: (i) identifying region of genomic DNA containing VNTR locus to get
unique electrophoretic pattern; (ii) repeating step (i) on similar sample of genomic DNA to obtain consistent
electrophoretic pattern; (iii) comparing the pattern obtained in step (ii) with different genomic DNA to
obtain unique band; and (iv) extracting the unique band of step (iii) to obtain said probe.
In yet another embodiment of the present invention, said equivalent probe is selected from a group comprising natural probe, synthetic probe and semi-synthetic probe or a combination thereof and can be obtained by plurality of ways selected from a group

comprising modification of one or more base of original probe, preferably replacing a base with inosine.
The present invention also relates to the use of nucleic acid probe or its equivalent probe for plurality of purposes selected from a group comprising consanguinity search, screening for tumoral disease, screening for hereditary disease and identification of biological specimen/cell line or a combination thereof
In still another embodiment of the present invention, said consanguinity search comprises steps of hybridization of said probe with nucleic acid of test individual/cell line, hybridization of said probe with nucleic acid of a second test individual/cell line and comparing size of hybridized fragments obtained from said hybridization to achieve consanguinity result.
In still another embodiment of the present invention, said screening for tumoral disease comprises steps of hybridization of said probe with nucleic acid of test cell of an individual/cell line, hybridization of said probe with nucleic acid of normal cell of same individual/cell line and comparing number and size of hybridized fragments obtained from said hybridization to achieve said screening for tumoral disease.
In still another embodiment of the present invention, said screening for hereditary disease comprises steps of hybridization of said probe with nucleic acid of test individual/cell line, hybridization of said probe with nucleic acid of heahhy individual/cell line and comparing number of hybridized fragments obtained from said hybridization to achieve said screening for hereditary disease.
In still another embodiment of the present invention, said identification of biological specimen/cell line comprises steps of hybridization of said probe with nucleic acid of biological specimen, hybridization of said probe with nucleic acid of test individual/cell line and comparing size of hybridized fragments obtained from said hybridization to achieve said screening for hereditary disease.
It has now been discovered that, using the DAMD technique, not only electrophoretic patterns can be established but also a signature pattern of bands hybridized to genomic DNA fragments (after hydrolysis with certain restriction enzymes) with a

specific fragment excised form the electrophoretic pattern could be established. The present invention relates to a method for selecting and/or obtaining nucleic acid probes capable of detecting VNTRs in the genome of a given species through a electrophoretic band pattern which might be unique in nature for a given species/cell line and the probe generated from these electrophoretic band further establish the pattern generated for a genome of given species by Southern analysis. The signature pattern by Southern analysis could also be unique for the genome of given species/cell line. According to the process of the invention, sets of primers for each locus of the VNTR regions designed and signature pattern are prepared for those primers of VNTR regions from the DNA of an individual /cell line. Selected unique bands are prepared as probes. These Probes are hybridized to the fragment of genomic DNA prepared by enzymatic hydrolysis of DNA from a genome bank of the species to which this individual belongs/ cell line, labeling these probes and placing them in contact, under hybridization conditions, with the restriction fragments of genomic DNA could establish a unique pattern of autoradiogram for a given genome by that particular probe.
Finally, selection is made of the probes (capable of hybridizing with said set of restriction fragments) which do not give hybridization profiles identical to those obtained with known probes and recognizing variable number tandem repeat regions.
Applications for the probes thus obtained include, particularly, processes for identifying an individual/cell line, Authenticity consanguinity testing, and investigating the origin of a seed.
In the present application, "probe" is understood to be any single-stranded nucleotide sequence that may match with a VNTR according to the well-known purine-pyrimidine matching properties of complementary nucleic acid strands in DNA-DNA, DNA-RNA, and RNA-RNA duplexes. This matching process is accomplished by the establishment of hydrogen bonds between the adenosine-thymine (A-T) and guanosine-cytosine (G-C) bases of double-stranded DNA; adenosine-uracil (A-U) base pairs may also form by hydrogen bonding in the DNA-RNA or RNA-RNA duplexes. Matching of nucleic acid strands to identify a nucleic acid molecule is usually called "nucleic acid hybridization" or simply "hybridization." Probes are with

the aid of at least one restriction enzyme cutting at an essentially constant distance from the site that it recognizes, then separation by size of the restriction fragments obtained, selection of fragments longer than 1.5 kb, and labeling said fragments by known methods, the thus obtained, labeled fragments of the same size constituting a probe.
"New VNTR" is understood to be a genome region not yet identified as such, for which no specific probe is known from the literature.
The present invention relates to a method for selecting and/or obtaining nucleic acid probes capable of detecting new VNTRs in the genome of a given species, characterized in that:
a) a regions of the genome containing these new sequence/existing VNTR loci identified by the bioinfo tool used to get the unique electrophoretic patterns using polymerase chain reaction (PCR) specific to these VNTR loci from the DNA of an individual of said species/cell line, said set being obtained, in a manner known of itself, by specific primers designed form new/exisiting VNTR loci and using the template genomic DNAcontaining at least part of the genomic DNA of said individual/cell line, with the aid of DNA polymerase,
b) the steps in paragraph a) above are repeated on similar samples of genomic DNA from the same individual/ cell line to obtain a consistant electrophoretic pattern sets,
c) in addition, the pattern generataed from step b) compared against the different individuals/ cell line a uniqueness of the pattern is established as a signature for that particular individual/cell line
d) each of the unique band identified from step c) will be gel extracted and used as probes against the genomic DNA prepared by enzymatic hydrolysis of DNA of a genomic bank of said species thus obtained probes is placed in contact with the digested genomic DNA, under hybridization conditions, a pattern of autoradiogram for each of the probes will be established.

e) a selection is made of the probes which, under these conditions, are capable of hybridization with restriction fragments, of at least a given length, from the set of fragments with which the probe in question was placed in contact. The signature pattern of autoradioagram of each probe and its uniquness pattern on different individuals/cell line will be assesed.
f) in addition, a selection is made of the probes which, with a set of restriction fragments obtained as in paragraph e) of DNA from an individual of the same species, do not give hybridization profiles identical to those obtained, with known probes recognizing VNTRs, on sets of restriction fragments obtained in a similar manner and coming from the same individual,
g) and, if desired, a nucleic acid probe including at least part of the sequence of the probes thus selected may be sequenced and/or synthesized and/or labeled with a tracer by known methods.
Of course it is possible, by using known methods, including those of paragraph e, f, g) above, to confirm that the nucleic acid probe obtained by the method of the invention does indeed recognize a VNTR. Further on in the specification, other methods for obtaining such a confirmation will be indicated.
The method according to the invention, it includes after step b) consisting of repeating steps d) then e) and f), challenged against the different sets of genomic DNA from individual/cell line. More over a different restriction eznyme used in step d) can lead to different pattern of autoradiogram as signatures obtained from the DNA of an individual /cell line. Of the probes capable of hybridizing with a set of restriction fragments coming from another individual/cell line, after step e'), those which give a hybridization profile different to that obtained with the fragments prepared in step c) are selected (in case the eletrophoretic pattern remains the same with two or more idividuals/cell line.
The restriction enzyme used in the method according to the invention is preferably an enzyme which recognizes a short and hence frequent sequence in the genome and has a restriction site at a constant or near-constant distance from the recognized site. Of

the enzymes possessing the required properties, one may cite in particular class II enzymes, namely those which cut at a specific site near the recognized site or included in the recognized site, and class III enzymes, which cut at a nonspecific site but at a constant or near-constant distance, on the order of 20 bases, from the recognized site. Of the usable enzymes, one may cite, for example, Alu I, Hae III, and Hinfl.
According to the method according to the invention, in step d) extraction of bands will be carried out with the well know estabalished methods and commerically avaiable gel extraction kits.
Separation of restriction fragments as a function of size may be accomplished by any known method, for example, by electrophoresis.
Of course, "fragment size" is understood to be fragment length, expressed as the number of nucleotides. The fragments are considered to be of the same size if, by using the separation process employed, said fragments are not separated. Thus, for example, when 1% agarose gel electrophoresis is used as the separation process, fragments are considered to be of the same size if their lengths differ by no more than approximately 50 to 100 bases.
"Genomic bank" designates any storage mode for the genome of an individual, which bank may be partial or total, namely containing all or part of said genome. The genome banks used are notably in the form of microorganisms into which, according to known methods, fragments of the genome or part of the genome to be stored are inserted. Among microorganisms in current use, bacteria, viruses, and yeasts in particular may be cited. The genome fragments to be stored are inserted directly into the genome of the recipient microorganism or are in an independent form, such as a plasmid. The use of these banks allows, through preculture of the microorganisms, the number of copies of the genome or the part of the genome to be studied to be easily increased. The sought-after genetic material is then extracted by known methods.

The probes used in the method according to the invention may be labeled by any marker in traditional use.
They may in particular be labeled using a radioactive tracer such as 32 P, 33 S, 125 H, and 14C. Radioactive labeling may be accomplished by known methods.
The nucleotide sequences may in particular be labeled at the 3' end by adding one or more deoxyribonucleotides or ribonucleotides, or a nucleotide analog such as a dideoxynucleotide, labeled at the alpha position by 32 P in the presence of deoxynucleotidyl terminal transferase; the nucleotide sequences can also be labeled at the 5' end with the aid of a kinase, for example T4 polynucleotide kinase; in this case, the radioactive phosphate is transferred from a nucleotide labeled at the gamma position to the polynucleotides. The nucleotide sequences can also be labeled at each end by adding any radiolabeled sequence in the presence of a ligase.
The probes can also be labeled by random priming or, during their chemical synthesis, by incorporation of one or more radioactive ribonucleotides or deoxyribonucleotides.
The hybridization detection method will depend on the radioactive label used and may be based on autoradiography, liquid scintillation, gamma radiation counting, or any technique allowing the radiation given off by the radioactive label to be detected.
Non-radioactive labeling may also be used, in a manner known of itself, for example by combining with the nucleotide sequences substances that have immunologic properties, such as an antigen or a hapten, or have a specific affinity for certain reagents, such as a ligand, or have properties allowing completion of enzymatic reactions such as an enzyme or an enzyme substrate. The nucleotide sequences can be labeled by random priming or, during their chemical synthesis, by incorporating one or more non-radioactively labeled ribonucleotides or deoxyribonucleotides. Non¬radioactive labeling can also be accomplished by incorporation into the 3' end of one or more deoxyribonucleotides or ribonucleotides, or a nucleotide analog such as a didesoxynucleotide including one of these groups. Labeling can also be done directly by chemical modification of the oligonucleotide, such as photobiotinylation or

sulfonation, or by fixation of DNA-specific dyes which are fluorescent when fixed such as the dimers of oxazole or thiazole orange. It can also be accomplished by addition of tracer molecules at the 3' or 5' end by chemical reaction after synthesis; also the nucleotide sequences can be labeled at each end by adding, in the presence of a ligase, any sequence including tracer molecules. The method of hybridization detection and development, carried out in a manner known of itself, will obviously depend on the non-radioactive label used.
The hybridization conditions of the method according to the invention are preferably discriminant conditions, thus allowing the specificity of said hybridization to be increased.
Among these conditions, one may cite in particular a temperature of between 50° and 65° C. and in particular between 65° C. and 70° C, and an ion concentration between 0.3 and 1.2M and preferably equal to 0.9M of Na + ions. The hybridization step proper is followed by several washings at 65° C. in a medium with a lower ion concentration, in particular between 0.015 and 0.15M of Na+^.
Using the method according to the invention, a hybridization profile is obtained for each set of fragments. When different hybridization profiles are obtained from sets of restriction fragments from one and the same individual placed in contact both with the probe obtained by the method and with known probes, it may be concluded that the probe obtained is new: namely, it is a probe able to identify a not-yet-known VNTR.
When different hybridization profiles are obtained from sets of restriction fragments from different individuals/cell line, placed in contact with one and the same probe, this confirms the variability of the VNTR detected. For this reason, the two individuals tested should preferably be unrelated in order to decrease the probability of revealing identical alleles.
When the hybridization profiles obtained are identical, namely when hybridization is observed on fragments of the same size, one may not conclude that the region detected lacks variability, as two hypotheses may then be considered: either the probe

used is indeed detecting a region that does not meet the definition of a VNTR, or the probe used is detecting a VNTR but the two individuals tested have identical alleles for the same region. To conclude, one may then analyze a set of restriction fragments coming from the DNA using different restriction enzymes obtained in the same manner as for the sets of fragments from the first two individuals/cell lines, and thus determine whether the sample tested is variable or not.
The present invention also relates to probes detecting new VNTRs that may be obtained by the method according to the invention.
Among the probes according to the invention, one may cite in particular:
The present invention also relates to synthetic or semi-synthetic polynucleotide probes equivalent to the new probes obtained by the method according to the invention (i.e. probes with the same hybridization properties). Modified probes may also be prepared, particularly ribonucleic acid probes or deoxyribonucleic acid probes in which one of the four bases is modified, and in particular, replaced by a base such as, for example, inosine.
The invention thus extends to any polynucleotide probe distinguished from a probe obtained directly by the method according to the invention only by the addition and/or deletion and/or substitution of one or more nucleotides while retaining the same properties of hybridization with the VNTR recognized by said probe directly obtained by said process. In particular, one may cite polynucleotides whose sequence is identical to that of a part of the probe initially obtained, but which is lacking the region or regions bordering the VNTR.
"Border region" is understood to be the 200 nucleotides located on either side of the VNTR proper.
The present invention also relates to the use of the probes described above.
The present invention relates in particular to the use of the probes as defined previously in consanguinity searches, screening for tumoral or hereditary diseases, or identification of a biological specimens/ cell lines etc.

The invention is further elaborated with the help of following examples; however, these examples should not be construed to limit the scope of the instant invention.
EXAMPLE 1;
HUMAN BEINGS/CELL LINE IDENTIFICATION/AUTHENTICATION; CONSANGUINITY SEARCH METHOD
According to one particular embodiment, the probes as defined above are used, particularly for human beings/cell line identification/authentication, in a consanguinity search method, comprising:
a first step in which said labeled probe or probes is/are each placed in contact, under hybridization conditions, with a set of restriction fragments obtained by hydrolyzing a sample of genomic DNA fi'om a tested individual/cell lines with the aid of at least one restriction enzyme cutting at an essentially constant distance from the site it recognizes, followed by separation of said fragments as a function of size;
a second step in which said labeled probe or probes is/are each placed in contact, under hybridization conditions, with a set of restriction fragments obtained by hydrolyzing a sample of genomic DNA from a second individual, whose consanguinity with said tested individual is to be determined, using the restriction enzyme or enzymes used in the first step, which fragments are also separated by size; and
a third step in which the sizes of the fragments hybridizing with said probe or probes are compared for the two individuals/cell line, it being understood that there is a presumption of consanguinity when the size of the fragments hybridizing with said probe or probes is identical.
One may conclude that consanguinity exists between two individuals/ cell line when a sufficiently representative number of fragments hybridizing with probes obtained according to the invention have the same size.

In particular, the process described above may be used for consanguinity testing by the method described by Alec Jeffreys ("highly variable minisatellites and DNA fingerprints", Biochemical Society Transactions, Vol. 15, pp. 309-317, 1987) or by Wong et al., ("Characterization of a panel of highly variable minisatellites cloned from Human DNA", Ann. Hum. Genet., 51, pp. 269-288, 1987).
This process may of course also be applied to verifying the origin of any animal or plant seed.
EXAMPLE 2;
SCREENING FOR TUMORAL DISEASES
The probes according to the invention can also be used for a process for screening genetic abnormalities associated with certain cancers or hereditary diseases.
The cancer-forming process is sometimes accompanied by general genetic instability bringing about mutations and/or losses of chromosome fragments, particularly fragments carrying antitumor genes. This general instability may result in a change in the number of pattern repeats of a VNTR or even in the deletion of a VNTR.
Hence the present invention relates to the use of probes as defined above in a screening process for tumoral diseases, characterized by comprising:
a first step in which said labeled probe or probes is/are each placed in contact, under hybridization conditions, with a set of restriction fragments obtained by hydrolyzing a sample of genomic DNA from test cells of an individual with the aid of at least one restriction enzyme cutting at an essentially constant distance from the site it recognizes, followed by separation of said fragments as a function of size;
a second step in which said labeled probe or probes is/are each placed in contact, under hybridization conditions, with a second set of restriction fragments obtained by hydrolyzing a sample of genomic DNA from normal cells of the same individual, using the restriction enzyme or enzymes used in the first step, which fragments are also separated by size; and

a third step in which a comparison is made, in both categories of cells, of the number and size of the fragments hybridizing with said probe or probes, it being understood that there is a presumption of tumoral disease when the number or size of fragments hybridizing with said probe(s) differs.
EXAMPLE 3:
SCREENING FOR HEREDITARY DISEASES
The probes as defined according to the invention can also be used in a process of screening for hereditary diseases in which losses of chromosome fragments carrying VNTR are observed, for example telomeric microdeletions associated with mental retardation.
The invention thus relates to the use of probes as defined above in a process of screening for hereditary diseases characterized by comprising:
a first step in which, under hybridization conditions, said labeled probe or probes is/are each placed in contact with a set of restriction fragments obtained by hydrolyzing a sample of genomic DNA fi-om a first individual to be tested, with the aid of at least one restriction enzyme cutting at an essentially constant distance from the site that it recognizes, said fragments then being separated by size,
a second step in which, under hybridization conditions, said labeled probe or probes is/are each placed in contact with a second set of restriction fragments obtained by hydrolyzing a sample of genomic DNA from a healthy individual, with the aid of the restriction enzyme or enzymes used previously, said fragments also being separated by size, and
a third step in which the number of fragments hybridizing with said probe or probes in the two individuals is compared, it being understood that there is a presumption of the existence of hereditary disease when the number of fi-agments hybridizing with said probe(s) differs.

Here, "healthy individual" is understood to be an individual who does not have the suspected hereditary disease.
EXAMPLE 4;
IDENTIFICATION OF A BIOLOGICAL SPECIMEN / CELL LINE
The probes according to the invention can also be used to determine whether a biological specimen comes from a given individual to be tested. This application is particularly useful in police investigations to determine whether a biological specimen indeed comes from an individual under suspicion.
According to this particular embodiment, the present invention thus relates to the use of probes as defined above in a process of identification of a biological specimen by comparison with the DNA of a test individual/cell line, characterized by comprising:
a first step in which, under hybridization conditions, said labeled probe or probes is/are each placed in contact with a set of restriction fragments obtained by hydrolyzing the DNA of said biological specimen, with the aid of at least one restriction enzyme cutting at an essentially constant distance from the site that it recognizes, followed by separation of said fragments as a fiinction of size, and autoradiogram pattern.
a second step in which, under hybridization conditions, said labeled probe or probes is/are each placed in contact with a second set of restriction fragments obtained by hydrolyzing a sample of genomic DNA from the individual tested, with the aid of the restriction enzyme or enzymes used in the first step, said fragments also being separated by size, and
a third step in which is compared the size of the fragments hybridizing with said probe(s) in the biological specimen and in the test individual/cell line, it being understood that there is a presumption that the biological specimen comes from the test individual when the size of the fragments hybridizing with said probe(s) is identical. Incase of identical status different restriction eznyme give a different

autoradiogram based on the size at which it hydrolysis the genomic DNA of the individual/cell line
The term "biological specimen" here designates any specimen from which DNA can be extracted to analyze it according to the process described above. This may in particular be a fragment of skin, saliva, sperm, etc.
The process described above also allows verification, in a similar manner, of whether the cells present in a biological specimen actually come from a given cell line.
REFERENCES:
1. Vergnaud, Gilles et al., "Detection of Single and Multiple Polymorphic Loci y Synthetic Tandem Repeats of Short Oligonucleotides," Electrophoresis, 12, pp. 134-140(1991).
2. Jeffreys, Alec. J., "Highly Variabel Minisatellites and DNA Fingerprints," Biochemical Society Transactions, vol. 15, Twenty-Third Colworth Medal Lecture, Sep. 1986, pp. 309-317.
3. Wong, Z. et al, "Characterization of a Panel of Highly Variable Minisatellites Cloned From Human DNA," Ann. Hum. Genet., 51, 269-288, (1987).
4. Zishcler, Hnas et al., "Dissecting (CAC).sub.5 /(GTG).sub.5 Multilocus Fingerprints From Man into Individual Locus-Specific, Hypervariable Components," Genomics 13, 983-990 (1992). 6. Armour, John A. L. et al., "Systematic Cloning of Human Minisatellites from Ordered Array Charomid Libraries," Genomics 8, 501-512 (1990).
5. Feinberg, Andrew P. et al., "A Technique for Radiolabeling DNA Restriction Endonuclease Fragments to High Specific Activity," Analytical Biochemistry, 132, 6-13 (1983).
6. Litt, M. et al., "A Highly Polymorphic Locus in Human DNA Revealed by Cosmid-Derived Probes," Proc. Natl. Acad. Sci., vol. 82, Sep. 1985, pp. 6206-6210.

7. Vergnaud, G. et al., "Detection, Cloning, and Distribution of Minisatellites in Some Mammalian Genomes," Experimentia Supplementum (Basel), DNA Fingerprinting: State of the Science; Ed. by S.D.J. Pena et al., vol. 67, pp. 47-57, (1993).
8. Nakamura et al., "Variable Number of Tandem Repeat (VNTR) Markers for Human Gene Mapping," Science, vol. 235, pp. 1616-1622, (1987).
9. AH et al., "Enzymatic Synthesis of DNA Probes Complementary to a Human Variable Number Tandem Repeat Locus", Analytical Biochem, 179, pp. 280-283,(1989).
10. Nakamura et al., "New Approach for Isolation of VNTR Markers", Am. J. Hum. Genet, vol. 43, pp. 854-859 (1988).

We Claim:
1) A nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR).
2) The nucleic acid probe as claimed in claim 1, wherein said equivalent probe is selected from a group comprising natural probe, synthetic probe and semi¬synthetic probe or a combination thereof and said modification comprises modification of one or more base, preferably replacing a base with inosine.
3) A method for obtaining nucleic acid probe or its equivalent probe capable of detecting Variable Number Tandem Repeat (VNTR), wherein said method comprises steps of:
(i) identifying region of genomic DNA containing VNTR locus to get
unique electrophoretic pattern; (ii) repeating step (i) on similar sample of genomic DNA to obtain
consistent electrophoretic pattern; (iii) comparing the pattern obtained in step (ii) with different genomic
DNA to obtain unique band; and (iv) extracting the unique band of step (iii) to obtain said probe.
4) The method as claimed in claim 3, wherein said equivalent probe is selected from a group comprising natural probe, synthetic probe and semi-synthetic probe or a combination thereof and can be obtained by plurality of ways selected from a group comprising modification of one or more base of probe of claim 1, preferably replacing a base with inosine.
5) Use of nucleic acid probe or its equivalent probe for plurality of purposes selected from a group comprising consanguinity search, screening for tumoral disease, screening for hereditary disease and identification of biological specimen/cell line or a combination thereof
6) The use of nucleic acid probe or its equivalent probe as claimed in claim 5, wherein said consanguinity search comprises steps of hybridization of said probe with nucleic acid of test individual/cell line, hybridization of said probe with

nucleic acid of a second test individual/cell line and comparing size of hybridized fragments obtained from said hybridization to achieve consanguinity result.
7) The use of nucleic acid probe or its equivalent probe as claimed in claim 5,
wherein said screening for tumoral disease comprises steps of hybridization of
said probe with nucleic acid of test cell of an individual/cell line, hybridization of
said probe with nucleic acid of normal cell of same individual/cell line and
comparing number and size of hybridized fragments obtained from said
hybridization to achieve said screening for tumoral disease.
8) The use of nucleic acid probe or its equivalent probe as claimed in claim 5,
wherein said screening for hereditary disease comprises steps of hybridization of
said probe with nucleic acid of test individual/cell line, hybridization of said
probe with nucleic acid of healthy individual/cell line and comparing number of
hybridized fragments obtained from said hybridization to achieve said screening
for hereditary disease.
9) The use of nucleic acid probe or its equivalent probe as claimed in claim 5,
wherein said identification of biological specimen/cell line comprises steps of
hybridization of said probe with nucleic acid of biological specimen,
hybridization of said probe with nucleic acid of test individual/cell line and
comparing size of hybridized fragments obtained from said hybridization to
achieve said screening for hereditary disease.
10) The nucleic acid probe or its equivalent probe, the method and the use as
substantially described herein with reference to examples.