高特异性聚合酶链反应法鉴别大、小黄鱼鱼鳔及其混淆品

目的:建立可鉴别大、小黄鱼鱼鳔及其混淆品的高特异性聚合酶链式反应(PCR)方法。方法:根据大、小黄鱼及6种常见混淆品线粒体12S r RNA基因序列,设计两对专用于大、小黄鱼鱼鳔的鉴别引物Fish-F1、Fish D-R1与Fish-F1、Fish X-R2;采用PCR法在不同的复性温度扩增,确立特异性反应条件。结果:在94℃预变性5 min;94℃变性30 s,60.4℃复性30 s,72℃延伸30 s,30个循环;72℃延伸5 min扩增条件下,大黄鱼鱼鳔得到约670 bp的扩增带,而小黄鱼和混淆品鱼鳔在同样的条件下无扩增带。在94℃预变性5 min;94℃变性30 s,64.8℃复性30 s,72℃延伸30 s,30个循环;72℃延伸5 min扩增条件下,小黄鱼鱼鳔得到约670 bp的扩增带,而大黄鱼和混淆品鱼鳔在同样的条件下无扩增带。结论:设计的鉴别引物对大、小黄鱼鱼鳔有高度的特异性。本方法简单、准确、快速、重现性好、稳定性高,能有效鉴别大、小黄鱼鱼鳔及其混淆品。

DIFFERENTIATION OF PSEUDOCONDYLOMA OF VULVA AND CONDYLOMA ACUMINATA BY DOT BLOT HYBRIDIZATION AND POLYMERASE CHAIN REACTION

This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82. 6%) out of 23 cases of condyloma acuminata and 2(25% ) out of 8 cases pseudocondyloma of vulvae (P<0. 05). PCR detected HPV DNA in 51 (92. 7%) out of 55 cases of condyloma acuminata , compared with none in 23 cases of pseudocondyloma (P<0.001 ). HPV DNA was present in the majority of condyloma acuminata specimens. HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection.