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Application of a PCR Method for the Detection of Mycoplasma in Veterinary Live Viral Vaccines  

Jeon Woo-Jin (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Kim Byoung-Han (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Jung Byeong-Yeal (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
An Dong-Jun (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Yi Chul-Hyun (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Jang Hwan (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Chung Gab-Soo (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
Publication Information
Korean Journal of Microbiology / v.41, no.4, 2005 , pp. 269-274 More about this Journal
Abstract
We evaluated the PCR assay and two commercialized PCR kits for the detection of mycoplasma in veterinary via live vaccines. The PCR assay could specifically detect all the tested Mycoplasma spp. and Acholeplasma spp., whereas two commercialized PCR kits did not. Also, the specificity of the PCR assay showed that 4 reference strains and 7 field isolates belonging to avian mycoplasma species could be all detected. The sensitivity of the PCR assay was determined using pure cultured Mycoplasma spp. and Acholeplasma spp. with a range of 1 to 100 colony forming units/ml in 9 CFR Mycoplasma broth. To test the availability of the PCR assay for veterinary live viral vaccines, A. laidlawii was artificially inoculated into the swine transmissible gastroenteritis-rota virus combined vaccine and canine parvovirus vaccine, respectively and the sensitivity of the PCR assay was similar with the result of cultured samples. In this study, the PCR assays could be used as rapid and sensitive methods for the detection of mycoplasma in veterinary live viral vaccines.
Keywords
mycoplasma detection; PCR; veterinary live viral vaccines;
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1 장명웅, 김광혁. 1993. 배양 세포주에서 Mycoplasmas의 오염검색. 대한미생물학회지 28, 209-221
2 한국동물약품협회. 1995. 국가검정동물용의약품검정기준. 1-9-18-06
3 Harasawa, R., H. Mizusawa., K. Nozawa., T. Nakagawa., K. Asada, and I. Kato. 1993. Detection and tentative identification of dominant Mycoplasma species in cell cultures by restriction analysis of the 16S-23S rRNA intergenic spacer regions. Res. Microbiol. 144, 489-493   DOI   ScienceOn
4 Hayflick, L. 1965. Tissue cultures and mycoplasmas. Tex. Rep. Biol. Med. 23, 285-303   PUBMED
5 Spaepen, M., A.F. Angulo., P. Marynen, and J.J. Cassiman. 1992. Detection of bacterial and mycoplasma contamination in cell cultures by polymerase chain reaction. FEMS Microbiol. Lett. 99, 89-94   DOI   ScienceOn
6 Tang, J., M. Hu., S. Lee, and R. Roblin. 2000. A polymerase chain reaction based method for detecting Mycoplasma/Acholeplasma contaminants in cell culture. J. Microbiol. Method. 39, 121-126   DOI   ScienceOn
7 Thornton, D.H. 1986. A survey of mycoplasma detection in veterinary vaccines. Vaccine 4, 237-240   DOI   PUBMED   ScienceOn
8 Toji, L.H., T.C. Lenchitz., V.A. Kwiatkowski., J.A. Sarama, and R.A. Mulivor. 1998. Validation of routine mycoplasma testing by PCR. In Vitro Cell. Dev. Biol. Anim. 34, 356-358   DOI   ScienceOn
9 Wirth, M., E. Berthold., M. Grashoff., H. PfUtzner., U. Schubert, and H. Hauser. 1994. Detection of mycoplasma contaminations by the polymerase chain reaction. Cytotechnology 16, 67-77   DOI   ScienceOn
10 Sasaki, T., R. Harasawa., M. Shintani., H. Fujiwara., Y. Sasaki., A. Horino., T. Kenri., K. Asada., I. Kato, and F. Chino. 1996. Application of PCR for detection of mycoplasma DNA and pestivirus RNA in human live viral vaccines. Biologicals. 24, 371-375   DOI   ScienceOn
11 Uphoff, C.C. and H.G. Drexler. 2002. Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines. In Vitro Cell. Dev. Biol. Anim. 38, 79-85   DOI   ScienceOn
12 Hu, M., C. Buck., D. Jacobs., G. Paulino, and H. Khouri. 1995. Application of PCR for detection and identification of mycoplasma contaminationin virus stocks. In Vitro Cell. Dev. Biol. Anim. 31, 710-715   DOI   ScienceOn
13 Benton, W.J., M.S. Cover, and F.W. Melchior. 1967. Mycoplasma gallisepticum in a commercial laryngotracheitis vaccine. Avian Dis. 11, 426-429   DOI   ScienceOn
14 Hay, R.J., M.L. Macy, and T.R. Chen. 1989. Mycoplasma infection of cultured cells. Nature. 339, 487-488   DOI   ScienceOn
15 Hopert, A., C.C. Uphoff., M. Wirth., H. Hauser, and H.G. Drexler. 1993. Specificity and sensitivity of polymerase chain reaction (PCR) in comparison with other methods for the detection of mycoplasma contamination in cell lines. J. Immunol. Methods. 164, 91-100   DOI   ScienceOn
16 전우진, 김병한, 정병열, 안동준, 이철현, 박수제, 주이석, 정갑수. 2004. 동물용 생물학적제제 및 세포배양에서 Mollicutes의 신속배양을 위한 배지의 선발. 한국마이코플라스마학회지 15, 28-34
17 Druckerei, C.H.B. 2002. Mycoplasmas, p.128-131. European Pharmacopoeia, 4th ed. Strasbourg press, Germany
18 Kobayashi, H., K. Yamamoto., M. Eguchi., M. Kubo., S. Nakagami., S. Wakisaka., M. Kaizuka, and H. Ishii. 1995. Rapid detection of mycoplasma contamination in cell cultures by enzymatic detection of polymerase chain reaction (PCR) products. J. Vet. Med. Sci. 57, 769-771   DOI   ScienceOn
19 Kojima, A., T. Takahashi., M. Kijima., Y. Ogikubo., M. Nishimura., S. Nishimura., R. Harasawa, and Y. Tamura. 1997. Detection of mycoplasma in avian live virus vaccines by polymerase chain reaction. Biologicals. 25, 365-371   DOI   ScienceOn
20 Wong-Lee, J.G. and M. Lovett. 1993. Rapid and sensitive PCR method for identification of Mycoplasma species in tissue culture, p. 257-260. In Persing D.H., T.F. Smith, F.C. Tenover, and T.J. White (eds.), Diagnostic molecular microbiology principles and applications, American Society for Microbiology, Washington, D.C
21 Raymond, A.M. 2002. Detection of mycoplasma contamination, p. 599-600. Code of Federal Regulations title 9, U.S. Government Printing Office Press, Washington
22 Barile, M.F., H.E. Hopps., M.W. Grabowski., D.B. Riggs, and R.A. DelGiudice. 1974. The identification and sources of mycoplasmas isolated from contaminated cell culture. Ann. NY. Acad. Sci. 225, 251-264
23 Stipkovits, L., L. Bodon., J. Romvary, and L. Varge. 1975. Direct isolation of mycoplasmas and acholeplasmas from sera and kidneys of calves. Acta. Microbiol. Acad. Sci. Hung. 22, 45-51   PUBMED
24 Teyssou, R., F. Poutiers., C. Saillard., O. Grau., F. Laigret., J.M. Bove, and C. Bebear. 1993. Detection of mollicute contamination in cell cultures by 16S rDNA amplification. Mol. Cell. Probes. 7, 209-216   DOI   ScienceOn
25 Razin, S., D. Yogev, and Y. Naot. 1998. Molecular biology and pathogenicity of mycoplasmas. Microbiol. Mol. Biol. Rev. 62, 1094-1156   PUBMED
26 Eldering, J.A., C. Felten., C.A. Veilleux, and B.J. Potts. 2004. Development of a PCR method for mycoplasma testing of Chinese hamster ovary cell cultures used in the manufacture of recombinant therapeutic proteins. Biologicals 32, 183-193   DOI   PUBMED   ScienceOn