COMPARISON OF DIFFERENT PRIMER SETS FOR DETECTION OF CHLAMYDIA-TRACHOMATIS BY THE POLYMERASE CHAIN-REACTION

被引:39
|
作者
ROOSENDAAL, R
WALBOOMERS, JMM
VELTMAN, OR
MELGERS, I
BURGER, C
BLEKER, OP
MACLAREN, DM
MEIJER, CJLM
VANDENBRULE, AJC
机构
[1] FREE UNIV AMSTERDAM,DEPT CLIN MICROBIOL,DE BOELELAAN 1117,1081 HV AMSTERDAM,NETHERLANDS
[2] FREE UNIV AMSTERDAM,DEPT PATHOL,1081 HV AMSTERDAM,NETHERLANDS
[3] ONZE LIEVE VROUW HOSP,DEPT GYNAECOL,1091 HA AMSTERDAM,NETHERLANDS
[4] FREE UNIV AMSTERDAM,DEPT PURE & APPL CHEM,1081 HV AMSTERDAM,NETHERLANDS
关键词
D O I
10.1099/00222615-38-6-426
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The sensitivity and specificity of the polymerase chain reaction (PCR) method was studied in vitro with HeLa cells infected with Chlamydia trachomatis serovar L2. Three different primer sets were studied; they were derived from the endogenous plasmid, the non-variable part of the MOMP gene and the 16S ribosomal RNA (rRNA) gene. The plasmid primers were the most sensitive in the PCR method and detected at least 0.1 infectious unit of C. trachomatis in the presence of a superfluous amount of human DNA. Application of this plasmid PCR to 13 C. trachomatis culture-positive cervical smears containing < 10-> 200 inclusion-forming units showed that it was the most sensitive of the three methods and detected C. trachomatis in all samples. This correlates with the observation that the plasmid PCR method could detect C. trachomatis in cervical smears of four symptomatic patients for up to 3 weeks after the start of treatment with doxycycline. In contrast, the MOMP gene- and rRNA gene-directed PCR, as well as culture and direct immunofluorescence, gave negative results within 1 week. Therefore, we conclude that the plasmid primers are the best candidates for use in the PCR method in C. trachomatis screening programmes and clinical follow-up studies.
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页码:426 / 433
页数:8
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