运用实时荧光PCR法辅助鉴定能力验证样品中的沙门氏菌

目的运用实时荧光PCR法辅助鉴定能力验证样品中的沙门氏菌,以提高检验准确度。方法采用传统的增菌、选择性培养、生化鉴定方法进行样品检验,实时荧光PCR法作为辅助手段对BPW增菌液及可疑菌落进行扩增,最后用API20E鉴定系统对可疑菌落进行确认。结果 3个样品经增菌、选择性培养、生化鉴定后均无显著符合沙门氏菌菌落特征及生化特征的可疑菌落;缓冲蛋白胨水增菌液实时荧光PCR法扩增结果表明样品S02为可疑样品,其分离得到的菌落S02-4有典型扩增曲线;经API20E鉴定系统确认,样品S02为阳性样品,阳性菌为猪霍乱沙门氏菌亚利桑那亚种。结论实时荧光PCR法操作简单,准确度高,可作为传统检验方法的有效补充。

Aided test the Salmonella in the samples of proficiency testing by real-time fluorescent PCR

Objective The real-time PCR was used to assist test the Salmonella in proficiency testing as to improve accuracy. Methods The samples were tested by traditional methods of enrichment, selective culture and biochemical identification. The real-time fluorescent PCR as an assistant method to amplify BPW enrichment solution and suspicious colonies. Finally, suspicious colonies were identified by API20E identification system. Results No suspicious colonies were found in the three samples after enrichment, selective culture and biochemical identification. Real-time fluorescent PCR amplification with BPW enrichment solution showed that sample S02 was suspicious and the colony S02-4 isolated from the sample S02 had typical amplification curve. Finally the API20E identification system confirmed that the sample S02 was positive and the suspicious bacteria were Salmonella cholerae subspecies Arizona. Conclusions The real-time PCR is simple to operate with high accuracy and can be used as an effective complement to the traditional testing methods.