TAILIEUCHUNG - Báo cáo y học: "Detection of poliovirus by ICC/qPCR in concentrated water samples has greater sensitivity and is less costly using BGM cells in suspension as compared to monolayers"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Detection of poliovirus by ICC/qPCR in concentrated water samples has greater sensitivity and is less costly using BGM cells in suspension as compared to monolayers | Balkin and Margolin Virology Journal 2010 7 282 http content 7 1 282 VIROLOGY JOURNAL SHORT REPORT Open Access Detection of poliovirus by ICC qPCR in concentrated water samples has greater sensitivity and is less costly using BGM cells in suspension as compared to monolayers Helene B Balkin Aaron B Margolin Abstract The integrated cell culture quantitative reverse transcriptase PCR ICC qRT-PCR method is used in our lab to detect enteroviruses in environmental waters. Typically we utilize monolayers of 3 cell lines buffalo green monkey kidney BGM human colonic carcinoma CACO-2 and African rhesus monkey kidney MA104 with the intent of providing one or more permissive hosts to a wide range of enteroviruses. In this study the BGM cell line was used to compare poliovirus infectivity in conventional monolayer cultures to BGM cells in suspensions. Propagated virus was subsequently amplified by qRT-PCR. Our PCR data showed lower cycle threshold Ct values in the suspensions which corresponded to a higher rate of infectivity than that observed in the monolayers. The difference in Ct values was determined statistically significant by One-way ANOVA . Infecting BGM cells in suspensions required less hands-on time less chance of contamination and was more cost effective than utilizing the conventional monolayer technique. Findings Viral infection is suspected in 50 of all acute gastrointestinal illness 1 with the public being at greatest risk acquiring infection through wastewaters that contaminate drinking water sources recreational waters and shellfish harvesting waters 2 . ICC qRT-PCR is a proven method for the rapid detection of infective enteroviruses in environmental waters 3 4 . With this technique viruses which are generally present in low numbers are propagated in monolayers of a host cell line which increases the PCR target. Little published research is available using cell culture systems other than monolayers to screen environmental samples 5

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