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A nested PCR-based method for detection of Streptococcus agalactiae 16S rRNA in milk and its application

Published online by Cambridge University Press:  12 February 2007

Jia Yu-Ping ,
Zhou Dong-Shun ,
Zhao Hong-Kun ,
Wan Ren-Zhong ,
Liu Wen-Qiang  and
Hu Jing-Dong
Jia Yu-Ping
Affiliation:
Preventive Veterinary Medicine of the Animal Science and Veterinary Medicine Department, Shandong Agricultural University, Taian 271018, China
Zhou Dong-Shun
Affiliation:
Center of Laboratory Animal Science, Shandong Academy of Medical Science, Jinan 250000, China
Zhao Hong-Kun*
Affiliation:
Preventive Veterinary Medicine of the Animal Science and Veterinary Medicine Department, Shandong Agricultural University, Taian 271018, China
Wan Ren-Zhong
Affiliation:
Preventive Veterinary Medicine of the Animal Science and Veterinary Medicine Department, Shandong Agricultural University, Taian 271018, China
Liu Wen-Qiang
Affiliation:
Preventive Veterinary Medicine of the Animal Science and Veterinary Medicine Department, Shandong Agricultural University, Taian 271018, China
Hu Jing-Dong
Affiliation:
Preventive Veterinary Medicine of the Animal Science and Veterinary Medicine Department, Shandong Agricultural University, Taian 271018, China
*
*Corresponding author. Email: hkzhao@sdau.edu.cn
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Abstract

Bovine mastitis caused by Streptococcus agalactiae is mainly subclinical and therefore can be diagnosed only in the laboratory. A nested polymerase chain reaction (PCR)-based method for specific, sensitive and rapid detection of S. agalactiae in raw milk was developed. The general streptococci primers, which anneal to conserved areas within the 16S rRNA subunit gene, were used as positive controls. The specificity of S. agalactiae primers is based on various areas within conserved areas of the 16S rRNA genes of S. agalactiae. Results have indicated that the method enables the detection of 1 CFU/ml of S. agalactiae in raw milk after enrichment, followed by DNA extraction using a rapid and simple procedure developed for this purpose, and specific PCR reaction. The method developed can be used efficiently in the early infectious status investigation of S. agalactiae in the dairy herd and in prevention and control of S. agalactiae spread in a herd.

Keywords

cow mastitisPCR16S rRNAStreptococcus agalactiae
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 3 , Issue 2 , August 2006 , pp. 115 - 118
Copyright
Copyright © China Agricultural University and Cambridge University Press 2006

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