We claim:
1. A method of performing Serum Bactericidal Activity (SBA) assay, the method comprising:
a. preparing a media and a buffer;
b. preparing a bacterial stock with a mother culture stock, the media and the buffer;
5 c. diluting the bacterial stock with the buffer to obtain a diluted bacterial stock;
d. adding a test serum, a complement, the diluted bacterial stock, and the buffer to
form an assay solution;
e. incubating the assay solution, spotting the assay solution on an agar plate and
incubating the agar plate; and
10 f. calculating antibody titre to determine the serum bactericidal activity.
2. The method of SBA assay as claimed in claim 1, wherein the media includes lysogeny
broth, Luria Bertani broth, Hi Soy (Hi Bacteriological media), tryptone, peptone, yeast
extract, or combinations thereof.
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3. The method of SBA assay as claimed in any one of claims 1 or 2, wherein the buffer
includes saline, phosphate, phosphate buffered saline, bovine serum albumin, Hanks'
balanced salt solution (HBSS), fetal bovine serum (FBS), or combinations thereof.
20 4. The method of SBA assay as claimed in any one of claims 1 to 3, wherein the buffer
includes HBSS, optionally comprising calcium and magnesium.
5. The method of SBA assay as claimed in any one of claims 1 to 3, wherein the buffer
includes HBSS and FBS.
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6. The method of SBA assay as claimed in any one of claims 1 to 5, wherein the agar plate is
Luria Bertani agar plate.
7. The method of SBA assay as claimed in any one of claims 1 to 6, wherein the bacterial
30 stock is prepared with bacteria in initial log phase, in exponential phase, in late Log phase,
or in a stationary growth phase;
optionally, wherein the bacterial stock preparation includes incubating bacteria from the
mother culture stock at 35? to 39? for 12 to 16 hours on a blood agar plate and
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optionally, wherein the bacterial stock preparation includes further incubating bacteria
from the blood agar plate on the