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Construction of a one-step multiplex real-time PCR assay for the detection of serogroups A, B, and E of associated with bovine pasteurellosis. | LitMetric

AI Article Synopsis

Article Abstract

Bovine pasteurellosis, caused by serogroups A, B, and E of (Pm), is mainly manifested as bovine respiratory disease (BRD) and hemorrhagic septicemia (HS). The disease has caused a great economic loss for the cattle industry globally. Therefore, identifying the Pm serogroups is critical for optimal diagnosis and subsequent clinical treatment and even epidemiological studies. In this study, a one-step multiplex real-time PCR assay was established. Three pairs of specific primers were prepared to detect the highly conserved genomic regions of serogroups A (), B (), and E () of Pm, respectively. The results depicted that the method had no cross-reaction with other bovine pathogens (, infectious bovine rhinotracheitis virus, and ). The linear range (10 to 10 copies/μL) showed the R values for serogroups A, B, and E of Pm as 0.9975, 0.9964, and 0.996, respectively. The multiplex real-time PCR efficiency was 90.30%, 90.72%, and 90.57% for CartA, CartB, and CartE, respectively. The sensitivity result showed that the serogroups A, B, and E of Pm could be detected to be as low as 10 copies/μL. The repeatability result clarified that an intra-assay and an inter-assay coefficient of variation of serogroups A, B, and E of Pm was < 2%. For the clinical samples, the detection rate was higher than the OIE-recommended ordinary PCR. Overall, the established one-step multiplex real-time PCR assay may be a valuable tool for the rapid and early detection of the serogroups A, B, and E of Pm with high specificity and sensitivity.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10336434PMC
http://dx.doi.org/10.3389/fvets.2023.1193162DOI Listing

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