{"title":"[多重PCR检测非o157 H7 STEC毒力基因的建立及应用]。","authors":"Xiao-Guang Wang, Ying-Hua Zhang, Ping Wang, Xiu-Hua Chen, Ling-Fei Luo, Yun Liu, Ji-Qian Liu, Chi-Ping Song, Yang Lin Ou, Guo-Qiang Chen","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>Traditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.</p><p><strong>Methods: </strong>Six virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.</p><p><strong>Results: </strong>The sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.</p><p><strong>Conclusion: </strong>The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.</p>","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 5","pages":"388-91"},"PeriodicalIF":0.0000,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection].\",\"authors\":\"Xiao-Guang Wang, Ying-Hua Zhang, Ping Wang, Xiu-Hua Chen, Ling-Fei Luo, Yun Liu, Ji-Qian Liu, Chi-Ping Song, Yang Lin Ou, Guo-Qiang Chen\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>Traditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.</p><p><strong>Methods: </strong>Six virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.</p><p><strong>Results: </strong>The sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.</p><p><strong>Conclusion: </strong>The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.</p>\",\"PeriodicalId\":70973,\"journal\":{\"name\":\"中华实验和临床病毒学杂志\",\"volume\":\"27 5\",\"pages\":\"388-91\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2013-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华实验和临床病毒学杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华实验和临床病毒学杂志","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的:传统的非o157产志贺毒素大肠杆菌检测方法在疾病监测中费时费力。基于多重PCR的毒力基因检测不仅提高了检测效率，而且提高了检测精度。方法:采用两组三倍pcr检测非o157:H7病毒的6个毒力基因(stx1、stx2、eae、hly、etpD、katP6)。采用SYBR Green I实时PCR的熔融曲线分析优化多重PCR。多重PCR检测结果与血清学检测结果一致。结果:多重PCR对stx1、stx2、eaeP、etpD、katP、hly的敏感性限分别为10 ng/ml、120 ng/ml、110 ng/ml、165 ng/ml、85 ng/ml、15 ng/ml，与单一PCR相似。对120例成人和90例儿童腹泻标本进行多重PCR检测，检出非o157阳性13例。结论:所建立的方法可用于非o157产志在大肠杆菌毒力基因的检测和筛选，具有较高的效率和准确性。
[Establishment and application of multiplex PCR for non-O157 H7 STEC virulence genes detection].
Objective: Traditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.
Methods: Six virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.
Results: The sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.
Conclusion: The method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.
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