单克隆抗体夹心酶标法检测鞭毛抗原快速诊断伤寒

Rapid diagnosis typhoid fever by detecting flagellin of salmonella typhi with McAb ELISA

目的建立一种早期、快速诊断伤寒的检验方法。方法用酸断裂法提取伤寒沙门菌鞭毛抗原免疫ＢＡＬＢ／ｃ鼠，用淋巴细胞杂交瘤技术制备得１Ｄ５、２Ｈ４、３Ａ６三株鞭毛抗原的单克隆抗体，以２Ｈ４、３Ａ６两株单抗标记辣根过氧化物酶，建立双抗体夹心酶联免疫吸附测定，用于定量检测血清中的可溶性伤寒沙门菌鞭毛抗原。结果检测灵敏度可达０．０７８ｎｇ／ｍｌ，批内及批间变异系数为４．９％和６．９％。检测血培养阳性的伤寒患者血清３４份，３２例阳性，符合率９４％。结论该方法可作为早期。

Objective To provide a rapid and early detecting measure for patients with typhoid fever. Methods BALB/c mice were immunized by purified flagellin from salmonella typhi according to Ibrahrim′s method. Four murine monoclonal antibodies against S. typhi flagellin were produced. Double antibodies ELISA was developed to detect flagellin of S. typhi in serum samples. Results This assay was able to detect as low as 0.078 ng/ml flagellin with interrun and inrun CV 6.9% and 4.9%. Using this assay, we detected S. typhi flagellin antigens in 94% (32/34) of serum samples from patients with positive hemoculture for S. typhi. Conclusion The method is of practical value in etiological diagnosis of typhoid fever.