FAST MULTIPLEX POLYMERASE CHAIN-REACTION ON BOILED CLINICAL-SAMPLES FOR RAPID VIRAL DIAGNOSIS

被引：49

作者：

VANDENVELDE, C ^{[1
]}

VERSTRAETE, M ^{[1
]}

VANBEERS, D ^{[1
]}

机构：

[1] BRUGMANN UNIV HOSP,DEPT VIROL,4 PL VAN GEHUCHTEN,B-1020 BRUSSELS,BELGIUM

来源：

JOURNAL OF VIROLOGICAL METHODS | 1990年 / 30卷 / 02期

关键词：

Fast Multiplex PCR; HPV infection; PCR optimization;

D O I：

10.1016/0166-0934(90)90022-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An assessment of optimal conditions for rapid simultaneous amplification of multiple human papillomavirus (HPV) sequences has been made using Thermus aquaticus DNA polymerase. All variables of practical value were studied by amplifying known target-sequences from ten-fold dilutions of well characterized cell lines. In our hands. Fast Multiplex PCR (FM-PCR), the technique of running multiple PCR reactions simultaneously with minimum incubation time at each temperature, was highly sensitive (amplification factor = 5 × 109 after 50 cycles), specific (100%) and reproducible (100%) for several microbiological applications. Diagnosis was generally obtained in less than 5 h after sampling. The results show that, after optimization of assay conditions, efficiency and specificity of Multiplex PCR depends exclusively on the primers design and concentration of the primers. © 1990.

引用

页码：215 / 227

页数：13

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