COMPARISON OF UREASE GENE PRIMER SEQUENCES FOR PCR-BASED AMPLIFICATION ASSAYS IN IDENTIFYING THE GASTRIC PATHOGEN HELICOBACTER-PYLORI

被引:16
作者
LOPEZ, CR
OWEN, RJ
BANATVALA, N
ABDI, Y
HARDIE, JM
DAVIES, GR
FELDMAN, R
机构
[1] LONDON HOSP,COLL MED,DEPT ORAL MICROBIOL,LONDON E1 2AD,ENGLAND
[2] LONDON HOSP,COLL MED,GASTROINTESTINAL SCI RES UNIT,LONDON E1 2AJ,ENGLAND
[3] LONDON HOSP,COLL MED,QMW,DEPT EPIDEMIOL & MED STAT,LONDON E1 4NS,ENGLAND
关键词
HELICOBACTER PYLORI; UREASE GENES; POLYMERASE CHAIN REACTION;
D O I
10.1006/mcpr.1993.1065
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Different genomic DNA samples and primer sequences were evaluated in urease (ure) gene-based PCR assays for rapid identification of Helicobacter pylori. Purified DNA and heated (boiled) cell lysates of bacterial cultures from gastric biopsies were tested with three primer sets for unique internal ureA, ureA + B and ureC sequences. The heated-lysates of H. pylori were quick to prepare but more frequently gave unexted variable or negative PCR results than assays performed on purified DNA, which were highly specific and reproducible for all three primer sets. Results indicated that sensitivity of the assay was linked to the size of the amplified target region rather than any particular strain feature, with the small 294 bp ureC product providing more accurate assays with heated-lysates of H. pylori. We strongly recommend that negative results in any PCR assay should be checked on purified DNA to exclude the possibility of a false-negative result. © 1993 Academic Press, Limited.
引用
收藏
页码:439 / 446
页数:8
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