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目的：建立幽门螺杆菌（ＨＰ）细胞空泡毒素（ＶＣ）的聚合酶链反应（ＰＣＲ）检测方法。方法：根据ＨＰＶＣ基因图谱，设计了两对顺序特异性引物，分别扩增包括信号肽顺序（４０６－１５１６）和不含此顺序（８１１－１５１６）的基因片段。对１４株经形态学和生化鉴定证实为ＨＰ的菌株提取其基因组ＤＮＡ，以上述引物进行ＰＣＲ。结果：已知ＶＣ阳性的ＨＰ标准株（ＮＣＴＣ１１６３８）、临床分离株ＮＺ７、ＮＺ８、ＨＰ２３、ＨＰ３５、ＨＰ５７、ＨＰ６５、ＨＰ６６、ＨＰ６７和ＨＰ７０共１０株为ＶＣ阳性。扩增产物中均检出ＶＣ特异的１１００ｂｐ、７０５ｂｐ、５４３ｂｐ片段，敏感性约为０．６ｎｇ基因组ＤＮＡ。结论：所设计的引物和ＰＣＲ检测方法正确，可应用于临床检测。
Objective: To establish a polymerase chain reaction (PCR) method for detection of vacuolar detoxification of Helicobacter pylori (HP) cells. Methods: According to HPVC gene map, two pairs of sequence specific primers were designed to amplify the gene fragments including signal peptide sequence (406-1516) and without this sequence (811-1516) respectively. Genomic DNA was extracted from 14 strains confirmed to be morphologically and biochemically identified as HP, and PCR was performed using the above primers. Results: VC positive were found in 10 VC positive HP standard strains (NCTC11638) and clinical isolates NZ7, NZ8, HP23, HP35, HP57, HP65, HP66, HP67 and HP70. The amplification products were detected VC specific 1100bp, 705bp, 543bp fragments, the sensitivity of about 0.6ng genomic DNA. Conclusion: The designed primers and PCR detection methods are correct and can be applied to clinical testing.