Detection of Mycoplasma species in cell culture by PCR and RFLP based method: effect of BM-cyclin to cure infections.
Indian J Med Microbiol
;
2007 Oct; 25(4): 364-8
Artigo
em Inglês
| IMSEAR
| ID: sea-53585
ABSTRACT
PURPOSE:
A two-stage nested polymerase chain reaction (PCR) assay system was described that amplifies the 16S-23S rRNA spacer region sequences of Mycoplasma and Acholeplasma infections in cell cultures and virus stocks.METHODS:
Established cell lines and virus stocks were screened for the presence of Mycoplasma by using nested PCR using two sets of outer and inner primers, amplifies 16S-23S rRNA. PCR and restriction fragment length polymorphism (RFLP) assay was used to detect and identify most of the species-specific Mycoplasmas involved in cell cultures and virus stock contaminants. Infected cultures detected by PCR-RFLP were further treated with BM-cyclin (5 microg/mL) and passaged for three times and tested for Mycoplasma infections by PCR-RFLP.RESULTS:
Mycoplasma pirum and Mycoplasma orale infections were detected by nested PCR. Species specificity was identified by using RFLP of Vsp I, Cla I and Hin dIII restriction enzymes. Mycoplasma infections were cured by treatment with BM-cyclin. This was further confirmed by non-amplification of PCR amplimers in BM-cyclin treated vs. non-treated cultures.CONCLUSIONS:
Regular monitoring of cell cultures for Mycoplasma infections and identification of species-specific Mollicutes will identify the source of contaminations. This approach can be used for quality control of the biological reagents used in cell culture and virology laboratories.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Controle de Qualidade
/
Virologia
/
Polimorfismo de Fragmento de Restrição
/
DNA Bacteriano
/
Reação em Cadeia da Polimerase
/
Técnicas de Cultura de Células
/
DNA Espaçador Ribossômico
/
Diterpenos
/
Minociclina
/
Antibacterianos
Tipo de estudo:
Estudo diagnóstico
Idioma:
Inglês
Revista:
Indian J Med Microbiol
Assunto da revista:
Microbiology
Ano de publicação:
2007
Tipo de documento:
Artigo
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