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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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References

Bo, RJ (1999) The antibacterial medication of bovine mastitis. Journal of Veterinary Medicine and Feed Additives 3: 14–16 (in Chinese with English abstract).Google Scholar
Chen, TS (1995) Manufacture and Application of the Microbial Culture Medium. Beijing: China Agricultural University Press.Google Scholar
Keefe, GP (1997) Streptococcus agalactiae mastitis: A review. Canadian Veterinary Journal–Revue Veterinaire Canadienne 38: 429–437.Google Scholar
Legal Sanitation and Supervision Department of Ministry of Health (2002) Technology and Criterion of Sterilization. Beijing: Ministry of Health, People's Republic of China.Google Scholar
Lenoard, J, Lascdea, SR and Dryyia, D (1984) Quanlitation of bacteria in cerebrospinal fluid and blood of children and its diagnostic significance. Journal of Clinical Microbiology 19(2): 187.Google Scholar
Li, HS, Yu, J, Luo, JY, Li, XP, et al. (2002) Bacteriological investigation of individual cattle farm mastitis in some regions (China). Journal of Traditional Chinese Veterinary Medicine 6: 14–17 (in Chinese with English abstract).Google Scholar
Liu, PH, Huang, Z, Lu, CP, Wang, J and Shen, LP (2004) Identification of bacteria causing cow mastitis and analysis of antibiotic resistance. Animal Husbandry and Veterinary Medicine 9: 13–15 (in Chinese with English abstract).Google Scholar
Shuang, J, Ga, ED, Bao, PY, Wu, YDL and Qi, ZL (2001) Occurrence regularity of dairy cattle's sub clinic mastitis: separation of pathogens and Medicine Sensitization Test. Journal of Inner Mongolia Agricultural University 22(1): 18–23 (in Chinese with English abstract).Google Scholar
Teng, K, Li, M, Yu, W, Li, H, Shen, D and Liu, D (1994) Comparison of PCR with culture for detection of Ureaplasma urealylicum in clinical samples from patients with urogenital infections. Journal of Clinical Microbiology 32(9): 2232–2234.CrossRefGoogle ScholarPubMed
William, C, Rebhun, DVM, Zhao, DM and Shen, JZ (1999) Diseases of dairy cattle [translation]. China Agricultural University Press 4: 384–385.Google Scholar
Zadoks, RN, González, RN, Boor, KJ and Schukken, YH (2004) Mastitis-causing streptococci are important contributors to bacterial counts in raw bulk tank milk. Journal of Food Protection 67(12): 2644–2650.CrossRefGoogle ScholarPubMed