Ratti G, Moroni A, Cevenini R
Sclavo Research Centre, Siena, Italy.
J Clin Pathol. 1991 Jul;44(7):564-8. doi: 10.1136/jcp.44.7.564.
A practical protocol using the polymerase chain reaction (PCR) was designed for detecting Chlamydia trachomatis in clinical samples. DNA was extracted from material collected on urethral swabs and used as substrate for the PCR. The target was a 600 basepair DNA segment of the multicopy plasmid that is common to all strains of the bacterium. Negative samples were checked for loss of DNA or presence of polymerase inhibitors by a second PCR, targeted to a conserved segment of the human genome. The whole procedure was tested on 216 men with non-gonococcal urethritis (NGU). All patients were independently assessed by tissue culture isolation (60 positive samples) and a commercial immunoenzymatic assay. The PCR protocol, while sufficiently simple for routine application, was reliable and, for the diagnosis of urethritis, at least as good as tissue culture isolation.
设计了一种使用聚合酶链反应(PCR)的实用方案,用于检测临床样本中的沙眼衣原体。从尿道拭子采集的样本中提取DNA,并用作PCR的模板。靶标是该细菌所有菌株共有的多拷贝质粒的一段600个碱基对的DNA片段。通过针对人类基因组保守片段的第二次PCR,检查阴性样本中DNA的丢失情况或聚合酶抑制剂的存在情况。对216例非淋菌性尿道炎(NGU)男性患者进行了整个检测过程的测试。所有患者均通过组织培养分离法(60份阳性样本)和一种商业免疫酶测定法进行独立评估。该PCR方案虽然简单到足以用于常规应用,但可靠,并且对于尿道炎的诊断,至少与组织培养分离法一样好。