利用稳定的新型铕荧光络合物BHHCT与Eu3+标记羊抗人HBsAb和Eu3+标记BSA-SA，建立定量测定血清HBsAb的Eu3+ -BHHCT-HBsAb-TRFIA法和Eu3+ -BHHCT-BSA-SA-TRFIA法。结果表明，这两种方法最低检出值分别为0.2ng/mL和0.05ng/mL，标准曲线范围均为0-100ng/mL，批内变异系数CV均小于10%后者回收率为85%-115%用Eu3+ -BHHCT-BSA-SA-TRFIA法与ELISA法同时检测118份血清样品，结果表明前者的检出率比后者高。

目的建立改良分子信标2双重实时PCR同时检测霍乱弧菌和副溶血弧菌的快速方法，应用于霍乱监测、副溶血弧菌食物中毒的快速诊断和海产品检验。方法根据GenBank公布的霍乱弧菌肠毒素基因A亚单位（ctxA）和副溶血弧菌的耐热直接溶血毒素基因（TDH）的保守序列，分别设计引物和改良分子信标探针，以10种细菌作对照，建立双重实时PCR2改良分子信标检测体系，应用于副溶血弧菌食物中毒快速诊断和霍乱监测。结果改良分子信标2双重实时PCR 反应体系DNA灵敏度为102.4～166.6fgPμl，菌液灵敏度为32～64CFUPml或3～6CFUPPCR反应体系，无交叉反应。此反应体系同时检测40株副溶血弧菌和50株霍乱弧菌，均出现特异的荧光信号，两种细菌检测互不干扰。对3起细菌性食物中毒共48份样品和100份海产品进行检测，9份副溶血弧菌实时PCR阳性，其中7份副溶血弧菌细菌培养阳性，其余样品都为阴性。从样品处理到检测结果仅需1天时间。结论改良分子信标2双重实时PCR 检测体系快速、灵敏度高、特异性强，可用于霍乱和副溶血弧菌食物中毒的快速诊断，为食源性疾病的分子流行病学调查提供新的检测手段。

以荧光染料Eam和Joe分别标记野生型和突变型双链探针作为均相检测探针，以构建的DNA模板作为研究模型 采用固定波长差同步荧光分析法对PCR反应产物进行终点检测，通过对HFE基因C282Y点突变的检测，并以限制性内切核酸酶Rsa1证实，该方法是一种廉价、快速、可靠的筛查遗传性血色病基因C282Y突变的方法，该法可扩展到各种基因的突变检测。

A rapid and simple homogeneous fluorescence PCR assay was developed for the clinical diagnosis of infectious diseases based on a molecular beacon. The established method could reproducibly detect Mycobacterium tuberculosis at the 10 bacteria/mL level. The analytical specificity was tested with 14 strains of mycobacterium, four unrelated bacteria and 220 negative samples; no false positive results were obtained. A blind test was also performed to evaluate its performance in Mycobacterium tuberculosis diagnosis. The results showed that both the clinical sensitivity and the specificity were 100%, and that the detection limit was in the range of 1-10 bacteria/ml. A clinical study with 466 patient samples demonstrated that fluorescence PCR assay correlated well with smear (93.6%) and culture (98.4%) methods for positive samples. However, fluorescence PCR could detect positive samples (62.9%) more than smear (30.3%) and culture (31.4%), indicating a higher sensitivity of the present method than the traditional ones. The feasibility of this method was further approved by successful detection of Neisseria gonorrhoeae and Chlamydia trachomatis.

目的建立改良分子信标-实时PCR检测霍乱弧菌的快速方法，应用于霍乱监测。方法根据GenBank公布的霍乱弧菌肠毒素基因A亚单位(ctxA)的保守序列，设计一对引物和改良分子信标探针，并进行特异性和灵敏度分析； 同时以11种细菌作对照，建立改良分子信标检测霍乱弧菌的实时PCR反应体系，应用于霍乱监测。结果霍乱弧菌改良分子信标检测体系检测12种细菌，只有霍乱弧菌有荧光信号，与其他细菌无交叉反应，DNA灵敏度为102.4fg/ul，菌液灵敏度为32cfu/ml或3cfu反应体系，对100份海产品和30份腹泻病人大便标本进行检测，结果都为阴性。检测时间仅需2h结论改良分子信标-实时PCR检测体系快速、灵敏度高、特异性强，可用于霍乱的快速诊断，为霍乱的分子流行病学调查提供新的检测手段。∀