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通过16S rRNA和gyrA基因与培养法比较,对AMPLICOR淋病奈瑟菌聚合酶链反应检测呈阳性的临床样本进行验证

Verification of clinical samples, positive in AMPLICOR Neisseria gonorrhoeae polymerase chain reaction, by 16S rRNA and gyrA compared with culture.

作者信息

Airell Asa, Lindbäck Emma, Ataker Ferda, Pörnull Kirsti Jalakas, Wretlind Bengt

机构信息

Department of Laboratory Medicine, Division of Clinical Bacteriology, F82 Karolinska University Hospital Huddinge, SE-141 86 Stockholm, Sweden.

出版信息

Int J STD AIDS. 2005 Jun;16(6):415-9. doi: 10.1258/0956462054094024.

Abstract

We compared 956 samples for AMPLICOR Neisseria gonorrhoeae polymerase chain reaction (PCR) (Roche) with species verification using the 16S rRNA gene to verification using gyrA gene. Control was the culture method. The gyrA verification uses pyrosequencing of the quinolone resistance-determining region of gyrA. Of 52 samples with optical density >/=0.2 in PCR, 27 were negative in culture, two samples from pharynx were false negative in culture and four samples from pharynx were false positives in verification with 16S rRNA. Twenty-five samples showed growth of gonococci, 18 of the corresponding PCR samples were verified by both methods; three urine samples were positive only in gyrA ; and one pharynx specimen was positive only in 16S rRNA. Three samples were lost. We conclude that AMPLICOR N. gonorrhoeae PCR with verification in gyrA gene can be considered as a diagnostic tool in populations with low prevalence of gonorrhoea and that pharynx specimens should not be analysed by PCR.

摘要

我们将956份用于淋病奈瑟菌聚合酶链反应(PCR)(罗氏公司)检测的样本,通过16S rRNA基因进行菌种鉴定与通过gyrA基因进行鉴定的结果进行了比较。对照方法为培养法。gyrA鉴定采用对gyrA喹诺酮耐药决定区进行焦磷酸测序的方法。在PCR检测中光密度≥0.2的52份样本中,27份培养结果为阴性,两份咽部分泌物样本培养结果为假阴性,四份咽部分泌物样本在16S rRNA鉴定中为假阳性。25份样本培养出淋病奈瑟菌,其中18份相应的PCR样本经两种方法鉴定均为阳性;三份尿液样本仅gyrA鉴定呈阳性;一份咽部分泌物标本仅16S rRNA鉴定呈阳性。三份样本丢失。我们得出结论,对于淋病低流行人群,采用gyrA基因鉴定的AMPLICOR淋病奈瑟菌PCR可被视为一种诊断工具,且咽部分泌物标本不应通过PCR进行分析。

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