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建立适用于临床检测血中伤寒沙门菌（Ｓ．ｔｙｐｈｉ）ＤＮＡ扩增方法。分离患者单核细胞，加热裂解制备模板ＤＮＡ，半巢式ＰＣＲ一步法扩增Ｓ．ｔｙｐｈｉ鞭毛基因。结果７例血培养阴性而临床拟诊伤寒及３８例由血培养阳性伤寒患者单核细胞均获得了４５８ｂｐ的扩增产物，其他２８例发热待查的患者单核细胞无扩增产物出现。连续１∶１０稀释Ｓ．ｔｙ－ｐｈｉＤＮＡ最低可检测到４０ｆｇＤＮＡ（约１０个细菌）。ｓｌｏｔ－ｂｌｏｔ探针杂交证实４５８ｂｐ扩增产物是Ｓ．ｔｙｐｈｉ鞭毛素基因。该法是一项简便、省时、灵敏和特异的实验室诊断伤寒的新技术，尤其对培养阴性的伤寒患者可提供早期、快速的实验室诊断。
To establish a DNA amplification method suitable for clinical detection of S. typhi in blood. The monocytes were isolated from the patients and heated for lysis to prepare the template DNA. Semi-nested PCR was used to amplify S. typhi flagella gene. Results A total of 458bp PCR products were obtained from 7 blood samples negative for clinical trial and 38 from mononuclear cells of blood culture positive typhoid fever. The other 28 patients with fever were not found in the mononuclear cells. S consecutive 1:10 dilution. The lowest detectable 40fgDNA of ty-phiDNA (about 10 bacteria). Slot-blot probe hybridization confirmed 458bp amplification product is S. typhi flagellin gene. The method is a simple, time-saving, sensitive and specific laboratory diagnosis of typhoid fever, especially in patients with negative typhoid fever can provide early, rapid laboratory diagnosis.