Pricila Khan Moly
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
A.F.M. Tariqul Islam
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
Md. Abdur Rakib
University of Science and Technology, Chittagong, Bangladesh
Md. Monirul Islam
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
Md. Golam Kabir
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
Pricila Khan Moly
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
A.F.M. Tariqul Islam
Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong-4331,
Md. Abdur Rakib
University of Science and Technology, Chittagong, Bangladesh
ABSTRACT
Studies were carried out on monoclonal antibodies (MAbs) ICL12, ICL26 and ICL31, which had been produced against Vibrio cholerae O1 and O139. These MAbs of IgG isotypes were used for the development of the Immunomagnetic Separation together with Polymerase Chain Reaction (IMS-PCR). Using MAbs ICL12, ICL26 and ICL31, the IMS-PCR procedure detected V. cholerae. in the pure cultures and in the stool samples. The detection limit of this method was 101 cfu mL-1 for V. cholerae. O139 while for the detection of V. cholerae. O1 Ogawa it was 102 cfu mL-1 and for V. cholerae. O1 Inaba it was 103 cfu mL-1. Of the 25 fresh stool samples tested, 15 were culture positive but 18 were IMS-PCR positive. Of the 55 frozen stool samples tested, 46 were culture positive but only 34 were IMS-PCR positive. However, when the 71 stool samples were frozen with Bovine Serum Albumin (BSA) added subsequently tested, 37 were culture positive while 49 were IMS-PCR positive. This showed that when stools were frozen with added BSA, the IMS-PCR procedure was much more specific and sensitive than the conventional culture method.
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How to cite this article
Pricila Khan Moly, A.F.M. Tariqul Islam, Md. Abdur Rakib, Md. Monirul Islam, Md. Golam Kabir, Pricila Khan Moly, A.F.M. Tariqul Islam and Md. Abdur Rakib, 2004. Detection of Vibrio cholerae O1 and O139 Serogroups Directly from Stool Specimens by Combined Immunomagnetic Separation and Polymerase Chain Reaction. Pakistan Journal of Biological Sciences, 7: 1654-1659.
DOI: 10.3923/pjbs.2004.1654.1659
URL: https://scialert.net/abstract/?doi=pjbs.2004.1654.1659
DOI: 10.3923/pjbs.2004.1654.1659
URL: https://scialert.net/abstract/?doi=pjbs.2004.1654.1659
REFERENCES
- Colwell, R.R., J.A.K. Hasan, A. Hug, L. Loomis and R.J. Siebling et al., 1992. Development and evaluation of a rapid, simple, sensitive monoclonal antibody based coagglutination test for the direct detection of Vibrio cholerae O1. FEMS Microbiol. Lett., 97: 215-219.
CrossRef - Mahbubur, R., D.A. Sack, S. Mahmood and A. Hossain, 1987. Rapid diagnosis of cholera by coagglutination test using 4-h fecal enrichment cultures. J. Clin. Microbiol., 25: 2204-2206.
Direct Link - Qadri, F., A. Chowdhury, J. Hossain, K. Chowdhury and T. Azim et al., 1994. Development and evaluation of rapid monoclonal antibody-based coagglutination test for direct detection of Vibrio cholerae O139 synony bengal in stool samples. J. Clin. Microbiol., 32: 1589-1590.
PubMed - Qadri, F., J.A.K. Hasan, J. Hossain, A. Chowdhury and Y.A. Begum et al., 1995. Evaluation of the monoclonal antibody-based kit bengal SMART for rapid detection of Vibrio cholerae O139 synonym bengal in Stool Samples. J. Clin. Microbiol., 33: 732-734.
Direct Link - Benenson, A.S., M.R. Islam and W.B. Greennough III, 1964. Rapid Identification of V. cholerae by dark field microscopy. Bull. WHO., 30: 827-831.
Direct Link - Bryton, P.R., M.L. Tamplin, A. Huq and R.R. Colwell, 1987. Enumeration of V. cholerae O1 in Bangladesh waters by fluorescent-antibody direct viable count. Appl. Environ. Microbiol., 53: 2862-2865.
PubMed - Bryton, P.R. and R.R. Colwell, 1986. Fluorescent antibody staining method for enumeration of viable environmental Vibrio cholerae O1. J. Microbiol. Methods, 6: 309-314.
CrossRef - Hasan, J.A.K., D. Bernstein, A. Huq, L. Loomis, M. Tamplin and R.R. Colwell, 1994. Cholera DFA: An improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of V. cholerae 01. FEMS Microbiol. Lett., 120: 143-148.
CrossRef - Sturbaum, G.D., P.T. Klonicki, M.M. Marshall, B.H. Jost, B.L. Clay and C.R. Sterling, 2000. Immunomagnetic separation (IMS)-fluorescent antibody detection and IMS-PCR detection of seeded Cryptosporidium parvum oocysts in natural waters and their limitations. J. Applied Environ. Microbiol., 68: 2991-2996.
CrossRef