两种核酸检测法在副溶血性弧菌耐热直接溶血素和不耐热溶血毒素中的应用评估  

Application of two nucleic acid detection methods in evaluation of thermostable direct hemolysin(TDH) and thermolabilehemolysin(TLH) of Vibrio parahaemolyticus

作　　者：黄世旺[1] 徐昌平[2] 方叶珍[1] 李剑[1] 赵雪琴[3] 李琳[4] 

机构地区：[1]杭州市江干区疾病预防控制中心,浙江杭州310004 [2]浙江省疾病预防控制中心,浙江杭州310051 [3]杭州市下城区疾病预防控制中心,浙江杭州310003 [4]杭州市上城区疾病预防控制中心,浙江杭州310009

出　　处：《疾病监测》2016年第9期746-749,共4页Disease Surveillance

基　　金：杭州市社会发展科研攻关项目(No.20130533B46);浙江省医药卫生科技计划(No.2014ZHA003)~~

摘　　要：目的比较CPA-核酸试纸条法与荧光定量PCR法检测副溶血性弧菌(VP)的tlh种特异性基因和tdh毒力基因的灵敏度与特异性。方法采用CPA-核酸试纸条法和荧光定量PCR法同时对655株不同来源的VP菌株进行tlh、tdh基因检测。结果 CPA-核酸试纸条法与荧光定量PCR法检测结果相同,VP菌株的tlh基因检测阳性率均为100%(655/655)。两种方法对tdh基因的检测结果也完全一致,tdh基因临床株与海产品株阳性检测率均分别为95.8%(207/216)与2.27%(10/439)。CPA-核酸试纸条法对tlh、tdh基因检测具有高度特异性,灵敏度可达6.2×103cfu/ml菌液,从核酸制备至完成检测最快仅需1 h。结论 VP临床株与涉水产品分离株普遍携带tlh种特异性基因,而tdh毒力基因主要存在于临床标本而在涉水产品株极少存在,可采用敏感、特异的CPA-核酸试纸条法,做VP腹泻患者的床边诊断以及食物中毒事故现场的快速检测。Objective To compare the sensitivity and specificity of CPA-nucleic acid test strip and real-time PCR for the detection of tlh specific genes and tdh virulence genes of Vibrio parahaemolyticus. Methods The tlh and tdh genes of 655 strains of V. parahaemolyticus w ere detected by using CPA-nucleic acid test strip and real-time PCR simultaneously. Results The results CPA-nucleic acid test strip and real-time PCR w ere consistent,the detection rates of tlh gene w ere all 100%（ 655 /655）. The detection results of the tdh gene w ere consistent too. The detection rates of the tdh gene isolated from clinical patients and seafood w ere 95. 8%（ 207 /216） and 2. 27%（ 10 /439） respectively. CPA-nucleic acid test strip had high specificity and sensitivity（ 6. 2 × 10^3 cfu / ml bacteria liquid） in the detections of tlh and tdh genes,only 1 hour w as needed to complete the test after the nucleic acid preparation. Conclusion V. parahaemolyticus strains isolated from clinical patients and seafood generally carry tlh specific gene,but tdh virulence gene mainly exist in clinical specimens. CPA-nucleic acid test strip can be used in the clinical diagnosis of diarrhea caused by V. parahaemolyticus and rapid field detection of food poisoning.

关 键 词：副溶血性弧菌 核酸检测法 评估 tlh TDH 

分 类 号：R516.3[医药卫生—内科学] R440[医药卫生—临床医学]