RT-PCR法在沙门菌检测及血清分型鉴定中的应用价值研究  

Study on the Application Value of RT-PCR in the Detection and Serotyping of Salmonella

作　　者：肖芳 周志强 刘冬梅 Xiao Fang(Laboratory Department of Yongxin County Center for Disease Control and Prevention,Yongxin Jiangxi 343400)

机构地区：[1]永新县疾病预防控制中心检验科,江西永新343400

出　　处：《黑龙江医药》2024年第3期692-696,共5页Heilongjiang Medicine journal

摘　　要：目的:探讨反转录-聚合酶链反应(RT-PCR)法在沙门菌检测及血清分型鉴定中的应用价值。方法:选择2022年8月至2024年1月本疾控中心收治的456例腹泻患者的粪便标本,均行细菌培养和RT-PCR检测,记录RT-PCR鉴定沙门菌血清分型结果,以细菌培养结果作为金标准,分析RT-PCR鉴定沙门菌的价值及其与细菌培养的一致性,对比细菌培养、RT-PCR鉴定沙门菌的费用及操作时间。结果:456例腹泻中经细菌培养共检出沙门菌78株,RT-PCR检出85株,RT-PCR检测敏感度为97.44%(76/78),特异度为96.83%(366/378),准确度为96.93%(442/456),阳性预测值为89.41%(76/85),阴性预测值为99.46%(366/368);RT-PCR鉴定沙门菌结果与细菌培养一致性较高(kappa值=0.897,P<0.001);RT-PCR鉴定结果与细菌培养完全一致有73株,符合率为93.59%,其中里森沙门菌、鼠伤寒沙门菌、德尔卑沙门菌、姆班达卡沙门菌4种优势血清型结果完全一致,而与细菌培养鉴定不一致菌株5株;RT-PCR鉴定沙门菌操作时间为(0.30±0.07)天,费用(100.22±6.52)元,细菌培养鉴定沙门菌的操作时间为(2.82±0.25)天,费用为(300.52±12.16)元,RT-PCR鉴定沙门菌的费用及操作时间均低于细菌培养,差异有统计学意义(P<0.001)。结论:RT-PCR法可准确检出沙门菌,判断沙门菌血清分型,与细菌培养结果具有较高的一致性,且操作用时短、所需费用低,可应用于沙门菌检测。Objective:To explore the application value of reverse transcription polymerase chain reaction(RT-PCR)in the detection and serotyping of Salmonella.Methods:456 stool samples of diarrhea patients admitted to our CDC from August 2022 to January 2024 were selected,and all of them were tested by bacterial culture and RT-PCR.The results of serotyping of Salmonella identified by RT-PCR were recorded.Taking the results of bacterial culture as the gold standard,the value of RT-PCR in identifying Salmonella and its consistency with bacterial culture were analyzed The cost and operation time of RT-PCR identification of Salmonella.Results:A total of 78 strains of Salmonella were detected in 456 cases of diarrhea by bacterial culture,85 strains were detected by RT-PCR,The sensitivity,specificity,accuracy,positive predictive value and negative predictive value of RT-PCR were 97.44%(76/78),96.83%(366/378),96.93%(442/456),89.41%(76/85)and 99.46%(366/368),respectively;The results of RT-PCR identification of Salmonella were highly consistent with the bacterial culture(kappa value=0.897,P<0.001),The results of RT-PCR identification were completely consistent with those of bacterial culture,with a coincidence rate of 93.59%.The results of four dominant serotypes of Salmonella reesen,Salmonella typhimurium,Salmonella delbe,and Salmonella Mbandaka were completely consistent,while five strains were inconsistent with those of bacterial culture identification;The operation time of RT-PCR to identify Salmonella was(0.30±0.07)days,and the cost was(100.22±6.52)yuan.The operation time of bacterial culture to identify Salmonella was(2.82±0.25)days,and the cost was(300.52±12.16)yuan,The cost and operation time of RT-PCR identification of Salmonella were lower than those of bacterial culture,and the difference was statistically significant(P<0.001).Conclusion:RT-PCR can accurately detect Salmonella and determine the serotype of Salmonella.It has high consistency with the results of bacterial culture,and has short operation time and low cost

关 键 词：沙门菌 反转录-聚合酶链反应 血清分型 细菌培养 

分 类 号：R446.5[医药卫生—诊断学] R378.22[医药卫生—临床医学]