Wong K C, Ho B S, Egglestone S I, Lewis W H
Department of Health Sciences, Hong Kong Polytechnic, Hung Hom, Kowloon.
J Clin Pathol. 1995 Feb;48(2):101-4. doi: 10.1136/jcp.48.2.101.
To evaluate the use of a duplex polymerase chain reaction (PCR) assay for the simultaneous detection of Neisseria gonorrhoeae and Chlamydia trachomatis in clinical samples.
Genital swab specimens were obtained from both China (203 swabs) and Hong Kong (202 swabs). N gonorrhoeae and C trachomatis were detected in each specimen with a number of tests including enzyme immunoassays (IDEIA) and PCR assays using both single and double primer pairs. The primer pair for N gonorrhoeae was derived from the cppB gene on its cryptic plasmid and the PCR product was 390 base pairs long. For C trachomatis, the PCR product was 473 base pairs long, resulting from amplification of a sequence in the common 7.4 kilobase plasmid present in all serovars. For N gonorrhoeae, PCR results were also compared with those obtained by culture and Gram's smear of the discharges.
For the 203 specimens collected in China, similar numbers of positive results (177) were obtained by both Gonozyme and duplex PCR for the detection of N gonorrhoeae. No discrepant results were found among the cultured specimens when Gonozyme and duplex PCR were compared. C trachomatis was detected in 47 specimens by duplex PCR, but was detected in only 28 by IDEIA. Of the 202 Hong Kong specimens, 46 were positive for N gonorrhoeae, detected by both Gonozyme and duplex PCR; 34 were positive for C trachomatis, 25 of which were detected by IDEIA and the remainder by duplex PCR.
The duplex PCR assay is a satisfactory diagnostic tool for the simultaneous detection of N gonorrhoeae and C trachomatis in clinical swab samples. Further evaluation is suggested.
评估双重聚合酶链反应(PCR)检测法在临床样本中同时检测淋病奈瑟菌和沙眼衣原体的应用。
分别从中国(203份拭子)和中国香港(202份拭子)获取生殖器拭子标本。采用多种检测方法对每份标本中的淋病奈瑟菌和沙眼衣原体进行检测,包括酶免疫分析(IDEIA)以及使用单引物对和双引物对的PCR检测法。淋病奈瑟菌的引物对源自其隐蔽质粒上的cppB基因,PCR产物长度为390个碱基对。对于沙眼衣原体,PCR产物长度为473个碱基对,是通过扩增所有血清型中常见的7.4千碱基质粒中的一个序列产生的。对于淋病奈瑟菌,PCR结果还与通过分泌物培养和革兰氏涂片获得的结果进行了比较。
在中国收集的203份标本中,Gonozyme法和双重PCR法检测淋病奈瑟菌的阳性结果数量相似(均为177例)。比较Gonozyme法和双重PCR法时,培养标本中未发现不一致的结果。双重PCR法检测出47份标本中有沙眼衣原体,而IDEIA法仅检测出28份。在中国香港收集的202份标本中,46份淋病奈瑟菌检测呈阳性,Gonozyme法和双重PCR法均检测到;34份沙眼衣原体检测呈阳性,其中25份通过IDEIA法检测到,其余通过双重PCR法检测到。
双重PCR检测法是在临床拭子样本中同时检测淋病奈瑟菌和沙眼衣原体的一种令人满意的诊断工具。建议进一步评估。