Development of a multiplex PCR method for detection of Pasteurella multocida and Mannheimia haemolytica
Vol. 132 No. 1C (2023), Original Article
Vol. 132 No. 1C (2023)

Development of a multiplex PCR method for detection of Pasteurella multocida and Mannheimia haemolytica

Original Article
https://doi.org/10.26459/hueunijns.v132i1C.7165
Published 2023-09-30
Nguyen Thi Thu Thao
Institute of Biotechnology, Hue University, Provincial Road 10, Phu Thuong, Hue, Vietnam
Nguyen Van Phu
Institute of Biotechnology, Hue University, Phu Thuong, Phu Vang, Thua Thien Hue, Vietnam
Nguyen Huu Tho
Institute of Biotechnology, Hue University, Provincial Road 10, Phu Thuong, Hue, Vietnam
Nguyen Thi Oanh
Institute of Biotechnology, Hue University, Provincial Road 10, Phu Thuong, Hue, Vietnam
Truong Phuc Hung
Thai Nguyen University of Sciences, Thai Nguyen University, Tan Thinh, Thai Nguyen, Vietnam
Ho Thi Xuan Tuy
Institute of Biotechnology, Hue University, Provincial Road 10, Phu Thuong, Hue, Vietnam
Nguyen Xuan Huy
Hue University, 3 Le Loi st., Hue, Vietnam
Le Cong Tuan
Hue University of Sciences, Hue University, 77 Nguyen Hue st., Hue, Vietnam
Nguyen Thi Thinh
Institute of Veterinary research and development of Central Vietnam, Km4, Vinh Hoa, Nha Trang, Vietnam
Nguyen Thi Kim Cuc*
Institute of Biotechnology, Hue University, Provincial Road 10, Phu Thuong, Hue, Vietnam
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How to Cite

1.
Nguyễn TTT, Nguyễn VP, Nguyễn HT, Nguyễn TO, Trương PH, Hồ TXT, Nguyễn XH, Lê CT, Nguyễn TT, Nguyễn TKC. Development of a multiplex PCR method for detection of Pasteurella multocida and Mannheimia haemolytica. hueuni-jns [Internet]. 2023Sep.30 [cited 2024Nov.23];132(1C):69-7. Available from: http://222.255.146.83/index.php/hujos-ns/article/view/7165
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Abstract

The multiplex PCR assay has been used to diagnose different diseases in livestock in Vietnam; however, this method has not been well-applied to sheep diseases. This study developed a multiplex PCR assay to detect the two main agents, Pasteurella multocida and Mannheimia haemolytica, causing Pasteurellosis in sheep. By using two pairs of specific primers, with the optimum concentration of each primer being 5 pM, this method successfully amplified the kmt1 gene from P. multocida and the gcp gene from M. haemolytica in the same reaction containing DNA of P. multocida and M. haemolytica at the lowest concentration of 0.1 ng each. Moreover, this multiplex PCR reaction is not affected by the artificial presence of some other bacterial DNA. In particular, this multiplex PCR showed 100% confidence in a stability test on a small scale with 20 raw DNA samples obtained from direct heat treatment or enrichment before heat treatment of 10 field samples. This is the first study in Vietnam on diagnosing P. multocida and M. haemolytica by multiplex PCR. This method is valuable for detecting P. multocida and M. haemolytica in sheep and serves as a reference for detecting these bacteria in other household animal.

https://doi.org/10.26459/hueunijns.v132i1C.7165
PDF (Vietnamese)

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