Methods for rapid identification of bacillus cereus

Details Methods for rapid identification of bacillus cereus Methods for rapid identification of bacillus cereus

2001-12-17

2004-03-02

Graser, Jennifer E. (Department: 1645)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

C435S007100, C435S007200, C435S007920, C435S034000, C435S975000

Reexamination Certificate

active

06699679

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to a method for rapid identification of
Bacillus cereus
and the kit thereof.
BACKGROUND OF THE INVENTION
Bacillis cereus
is gram-positive, spore-forming, motile, aerobic rod which inhibits in soil and has been recognized as an opportunistic food poisoning pathogen.
Bacillis cereus
and the poison thereof usually result in the diarrheal syndrome and the emetic syndrome. Some food types that are preferentially contaminated with
B. cereus
are crude cereals, starchy foods, dairy products, meat, dehydrated foods, and spices.
Conventional procedures for the detection of
B. cereus
are based on morphologic observations, physiological and biological tests, such as plate count method and most probable number method. Harmon et al. utilized the hydrolysis of lecithin (egg yolk reaction) as a characteristic for isolation of suspect
B. cereus
on selection media such as mannitol-egg yolk-polymyxin (MYP) agar. However, it should take several days to complete the physiological and biological tests. (Harmon, S. M. et al., 1992, Compendium of methods for the Microbiological examination of foods. 3
rd
ed. American Public Health Association, Washington, D.C., pages 593-604) Schraft et al. developed polymerase chain reaction for detection of lecithin-hydrolyzing gene of
B. cereus
. Since most strains of the
B. cereus
group (i.e.,
B. cereus, B. mycoides
and
B. thuringiensis
) possess lecithinase activity, the polymerase chain reaction method detects all species of the
B. cereus
group, but does not distinguish any single species. (Schraft, H. and M. W. Griffiths, 1995, Sepecific oligonucleotide primers for detection of lecithinase-positive Bacillus spp. by PCR. Appl. Environ. Microbiol. 61: 98-102). Thus, a rapid and specific method for identification of
B. cereus
is desired.
SUMMARY OF THE INVENTION
The objective of this invention relates to provide a method for rapid identification of
Bacillus cereus
in a sample comprising
(a) mixing an antibody or antisera specifically against the cell surface antigens of
B. cereus
with the sample, wherein the surface antigen is selected from the group of consisting of: surface antigens of
B. cereus
with molecular masses of 28.5, 26.5 and 20 kDa and the mixture thereof; and
(b) detecting the existence of
B. cereus
if the antibody-antigen binding reaction is positive.
Another objective of this invention is to provide a kit for rapid identification of
B. cereus
, which comprises a solution of antibody or antisera against the cell surface antigens of
B. cereus
, and the agents and apparatus required for detection of binding reaction between the said antibody or antisera and the bacteria antigen in a test sample; wherein the antigen is selected by at least one from the group consisting of: surface antigen of
B. cereus
with molecular masses of 28.5, 26.5 and 20 kDa and the mixture thereof.


REFERENCES:
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patent: 6284517 (2001-09-01), Restaino
patent: 2002/0082386 (2002-06-01), Mangold et al.
patent: 8602363 (1986-04-01), None
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Koo et al. May 19-23, 1996. ASM Abstracts. New Orleans, USA. 96(O). p. 381.*
Nielsen et al. J.Biol. Chem. 1982. 257(8): 4490-4495.*
Harmon et al. 1992. Compendium of methods for the Microbiological Examination of foods. 3rd ed. American Public Health Association. Washington DC., pp. 593-604. reference not included. Cited on p. 1 of Applicants′ specification.

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