Deguchi T, Yamamoto H, Iwata H, Ito Y, Ban Y, Tamaki M, Maeda S, Saito I, Ezaki T, Kawada Y
Department of Urology, Gifu University School of Medicine.
Kansenshogaku Zasshi. 1991 Dec;65(12):1555-9. doi: 10.11150/kansenshogakuzasshi1970.65.1555.
A polymerase chain reaction (PCR) with Chlamydia trachomatis-specific primers was applied for detection of C. trachomatis from urethral swab in male urethritis. The results were compared with those of culture method for detection of C. trachomatis. Of 18 clinical specimens tested in this study, inclusion bodies of C. trachomatis were detected in 11 specimens by the culture method. For PCR, sample DNA was prepared from transport medium in which urethral smear was suspended and two oligonucleotides based on sequences within the major outer membrane protein gene from C. trachomatis serovar L2 were used as extension primers. In 12 of the 18 specimens, 242bp DNA fragment was amplified by PCR and demonstrated to be the DNA fragment of C. trachomatis by Southern blot hybridization. No DNA of 242bp was amplified by PCR from five specimens in which any inclusion bodies of C. trachomatis were observed or from a specimen in which one inclusion body per cover slip was detected by culture method. C. trachomatis DNA of 242bp was amplified from all specimens in which 14 and more inclusion bodies per cover slip were detected by culture method. In two specimens concluded s negative by culture method, amplified C. trachomatis DNA were detected by PCR. Thus, the PCR would be a more simple and sensitive method for detection of C. trachomatis, compared with the culture method.
采用沙眼衣原体特异性引物的聚合酶链反应(PCR)检测男性尿道炎患者尿道拭子中的沙眼衣原体。将结果与沙眼衣原体培养检测方法的结果进行比较。在本研究检测的18份临床标本中,通过培养法在11份标本中检测到沙眼衣原体包涵体。对于PCR,从悬浮有尿道涂片的转运培养基中制备样本DNA,并使用基于沙眼衣原体血清型L2主要外膜蛋白基因序列的两种寡核苷酸作为延伸引物。在18份标本中的12份中,PCR扩增出242bp的DNA片段,通过Southern印迹杂交证明其为沙眼衣原体的DNA片段。在5份未观察到任何沙眼衣原体包涵体的标本或1份通过培养法检测到每盖玻片1个包涵体的标本中,PCR未扩增出242bp的DNA。在所有通过培养法检测到每盖玻片有14个及以上包涵体的标本中均扩增出242bp的沙眼衣原体DNA。在两份培养法判定为阴性的标本中,通过PCR检测到扩增的沙眼衣原体DNA。因此,与培养法相比,PCR将是一种更简单、灵敏的沙眼衣原体检测方法。