Development of a differential Multiplex PCR assay for Chlamydia abortus , Brucella spp. & Campylobacter fetus

BACKGROUND AND ABJECTIVEAbortion is one of the most critical factors affecting lambing rates. Chlamydia abortus, Brucella spp. and Campylobacter fetus are main pathogens because of the potential impact on veterinary and human health. Accurate, rapid, and specific methods are required to identify pathogens for controlling bacteria causing abortion in sheep.MATERIALS AND METHODSMultiplex PCR method was optimized for identification of bacterial infections caused by Chlamydia abortus , Brucella spp., Campylobacter fetus. The sensitivity and specificity of the test were evaluated. The number of ۵۷ samples of aborted sheep fetuses along with the placenta sent to the veterinary laboratory of North Khorasan province in ۲۰۲۱ was examined. The expected sizes of amplicons were ۲۲۲ bp for Brucella spp.( Baily et al., ۱۹۹۲), ۴۷۹ bp for Chlamydia abortus (DeGraves et al., ۲۰۰۳) and ۳۵۹ bp for Campylobacter fetus(Yamazaki et al., ۲۰۰۷).RESULTS AND DISCUSSIONIn this study, ۵۷ samples were analyzed by single and multiplex PCR method. Chlamydia abortus was detected in ۴۶ samples(۸۰%) that simultaneous presence of Chlamydia abortus and Brucella spp. was confirmed that in one samples. Brucella spp. alone was observed in ۵ samples, Therefore, in total, ۶ samples(۱۰%) were infected with Brucella spp., Campylobacter fetus was observed in ۳ sample(۵%).CONCLUSIONThis study confirmed the high prevalence of C.abortus, Brucella spp. and Campylobacter fetus among sheep and goat flocks in Iran. Diagnosis of infection by mPCR helps us to use appropriate antimicrobial agents to treat and control the disease. This technique is helpful in epidemiological studies for determinant focality of diseases and prevention of outbreaks. Multiplex PCR is a useful, inexpensive, accurate, and rapid method for identifying bacterial agents and more samples can be identified by this assay in a shorter period of time.

کلیدواژه ها:

Multiplex PCR ، Chlamydia abortus ، Brucella ، Campylobacter fetus