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Development of rapid PCR method for simultaneous identification of species, specific capsular type, and toxigenicity of Pasteurella sp. isolates
Authors:Ilnaz Sahragard  Yahya Tahamtan  Majid Valadan  Masoumeh Hyati  Farid Moazeni  Zahra Shirazi
Affiliation:1. Department of Microbiology, Islamic Azad University, Jahrom Branch, Jahrom, Iran
2. Department of Microbiology, Razi Vaccines and Serum Research Institute-Shiraz, P.O. Box 71955-367, Shiraz, Iran
Abstract:Toxigenicity, species, and type species of Pasteurella multocida isolates cannot be differentiated by morphology or standard biochemical reactions. A more rapid method is needed for P. multocida detection from clinical cases. These findings provide rapid insights into the characteristics of P. multocida isolates and suggest that this method can identify toxigenic and specific capsular type P. multocida. A PCR assay has been developed for rapid detection of P. multocida and differentiation of capsular types A and D. In this rapid method, kmt1, capA and capD, and toxA genes were amplified and a reliable multiplex PCR method for the detection of P. multocida in sheep and goats in the south of Iran was designed. Twenty isolates were obtained, which evinced characteristic morphological and cultural properties. Ten samples were identified simultaneously through the presence of the kmt1 gene as P. multocida species, the hydD–hydC gene as type A capsule, and the toxA gene as dermonecrotic toxin by mPCR, but none of them belonged to type D.
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