摘要
目的建立和评价一个新的多重和巢式PCR反向线点杂交试验(RLB)检测泌尿生殖道沙眼衣原体及其分型方法。方法该试验设计了3对引物分别针对CT外膜蛋白(omp1)基因VD2区和质粒进行扩增,用CT15种血清型(A、B/Ba、C、D/Da、E、F、G/Ga、H、I/Ia、J、Ja、K、L1、L2和L3)探针与扩增后的产物杂交。经COBASAmplicor检测的355份标本,其中277份尿液,2份男性直肠拭子和78份女性宫颈试子,进行PCRRLB检测CT并分型。结果192份COBASAmplicor阳性标本中175份PCRRLB阳性,其中93.1%(163/175)的CT感染可被正确分型,其单一型感染分型结果与DNA测序结果一致。163份COBASAmplicor阴性标本中6份巢式PCRRLB阳性。85.3%(139/163)为单一型CT感染,最常见的CT是E(38.6%)F(28.5%)和D(18.2%)。14.7%(24/163)为CT混合感染,混合感染以H和K为主。结论PCRRLB检测CT是一种简便、快速、特异和敏感的方法,并准确的进行CT分型,为CT感染的临床诊断和分子流行病学研究提供了一种可靠的方法。
Objective To develope and evaluate a new multiple and nested PCR-reverse line blot （RLB） hybridization assay for the detection of Chlamydia trachomatis infections and typing. Methods Twosets of primers directed to the VD2 region of the ompl gene of C. trachomatis and one set of primers directedto the cryptic plasmid of C. trachomatis were designed. The assay was developed for the detection and typingof 15 different serovars of C. trachomatis（ A、B/Ba,C,D/Da,E,F,G/Ga、H、I/Ia,J,Ja, K,L1、L2 and L3 ）.The assay developed was tested with 277 urine , rectal swab of 2 men and cervical scrapes of 78 women previously for C. trachomatis by using COBAS Amplicor test. Results 175 of 192 COBAS Amplicor-positivespecimens were positive by PCR- RLB. Using this assay, we were able to type 163 of the 175 samples（93. 1% ）. Six of 163 COBAS Amplicor -negative specimens were positive by nested PCR- RLB. Of thesesamples, 85.3% were single C. trachomatis infections, and 14. 7% were multiple infections. The mostcommon serovars identified were serovars E （ 38. 6% ）, F （ 28.5% ）, D （ 18.2% ） . Of the people withmultiple C. trachomatis infections contained main both serovars H and K, Conclusions The multiple PCR-RLB combine with nested PCR developed is a simple, fast, sensitive and specific method for the detectionand subtype of C. trachomatis. Moreover, it is an appropriate method for studying multiple C. trachomatisinfections and for use in large epidemiological studies.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2006年第3期251-253,共3页
Chinese Journal of Laboratory Medicine
基金
深圳市科研基金(20050116)
关键词
聚合酶链反应
核酸杂交
衣原体
沙眼
血清分型
Polymerase china reaction
Nucleic acid hybridization
Chlamydia trachomatis
Serotyping
