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Pakistan Journal of Biological Sciences
Year: 2004  |  Volume: 7  |  Issue: 10  |  Page No.: 1654 - 1659

Detection of Vibrio cholerae O1 and O139 Serogroups Directly from Stool Specimens by Combined Immunomagnetic Separation and Polymerase Chain Reaction

Md. Monirul Islam, Md. Golam Kabir, Pricila Khan Moly, A.F.M. Tariqul Islam and Md. Abdur Rakib    

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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