Rodkvamtook Wuttikon, Zhang Zhiwen, Chao Chien-Chung, Huber Erin, Bodhidatta Dharadhida, Gaywee Jariyanart, Grieco John, Sirisopana Narongrid, Kityapan Manerat, Lewis Michael, Ching Wei-Mei
Armed Forces Research Institute of Medical Science (AFRIMS), Royal Thai Army, Bangkok, Thailand; Naval Medical Research Center (NMRC), Silver Spring, Maryland; Preventive Medicine and Biometrics Department, Uniformed Services University of the Health Sciences (USUHS), Bethesda, Maryland.
Armed Forces Research Institute of Medical Science (AFRIMS), Royal Thai Army, Bangkok, Thailand; Naval Medical Research Center (NMRC), Silver Spring, Maryland; Preventive Medicine and Biometrics Department, Uniformed Services University of the Health Sciences (USUHS), Bethesda, Maryland
Am J Trop Med Hyg. 2015 May;92(5):967-71. doi: 10.4269/ajtmh.14-0627. Epub 2015 Mar 23.
We developed a rapid dot-enzyme-linked immunosorbent assay (dot-ELISA) using the combination of recombinant 56-kDa protein antigens that exhibited broad reactivity with serum antibodies against the four most prevalent strains (Karp, Kato, Gilliam, and TA763) of Orientia tsutsugamushi. The assay is rapid (30 minutes), and can be done at room temperature, and results can be read by the naked eye. Only a simple shaker is required to wash the membrane. Sera from 338 patients suspected of being ill with scrub typhus from rural hospitals around Thailand were tested using this dot-ELISA. Seventy-five (22.2%) patients were found to be positive. The sensitivity and specificity of dot-ELISA were determined using the indirect immunofluorescent assay (IFA) test as the gold standard, with the cutoff titer of immunoglobulin peroxidase conjugate M (IgM)/G (IgG) greater than 1:400/1:400. The dot-ELISA had a sensitivity of 98.5%, a specificity of 96.3%, a positive predictive value of 86.7%, and a negative predictive value of 99.6% for the acute-phase specimens. The results indicate that dot-ELISA rapid test using recombinant 56-kDa protein antigen was comparable with the IFA test and may be very useful for the diagnosis of scrub typhus in rural hospitals, where IFA is not available.
我们开发了一种快速斑点酶联免疫吸附测定法（斑点酶联免疫吸附测定），该方法使用重组56 kDa蛋白抗原的组合，该抗原与针对恙虫病东方体四种最常见菌株（Karp、Kato、Gilliam和TA763）的血清抗体具有广泛反应性。该测定法快速（30分钟），可在室温下进行，结果可用肉眼读取。仅需一个简单的振荡器来洗涤膜。使用这种斑点酶联免疫吸附测定法对来自泰国各地农村医院的338例疑似患恙虫病的患者血清进行了检测。发现75例（22.2%）患者呈阳性。以间接免疫荧光测定法（IFA）作为金标准，免疫球蛋白过氧化物酶结合物M（IgM）/G（IgG）的截断滴度大于1:400/1:400，来确定斑点酶联免疫吸附测定法的敏感性和特异性。对于急性期标本，斑点酶联免疫吸附测定法的敏感性为98.5%，特异性为96.3%，阳性预测值为86.7%，阴性预测值为99.6%。结果表明，使用重组56 kDa蛋白抗原的斑点酶联免疫吸附测定快速试验与IFA试验相当，对于没有IFA检测的农村医院诊断恙虫病可能非常有用。
