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酶免疫测定法检测隐孢子虫病患者纵向血清样本中的抗原特异性免疫球蛋白G抗体。

Enzyme immunoassay detection of antigen-specific immunoglobulin g antibodies in longitudinal serum samples from patients with cryptosporidiosis.

作者信息

Priest J W, Li A, Khan M, Arrowood M J, Lammie P J, Ong C S, Roberts J M, Isaac-Renton J

机构信息

Division of Parasitic Diseases, Centers for Disease Control and Prevention, Mail Stop F-13, Building 23, Room 1025, 4770 Buford Highway N.E., Atlanta, Georgia 30341-3724, USA.

出版信息

Clin Diagn Lab Immunol. 2001 Mar;8(2):415-23. doi: 10.1128/CDLI.8.2.415-423.2001.

Abstract

Cryptosporidium parvum is a protozoan parasite that causes diarrheal illness in a wide range of mammalian hosts, including humans. Characteristic serum immunoglobulin G (IgG) antibody responses to antigens in the 27- and 17-kDa size ranges have been shown to develop after infection, and several enzyme-linked immunosorbent assay (ELISA) and Western blot assay formats have been used to measure these IgG levels in human serum. Using a collection of serial samples from laboratory-confirmed cryptosporidiosis patients, we compared the results obtained by using two new ELISAs with those obtained with two different Western blot assays. When assayed with the large-format Western blot, 97% of the 67 patients had a demonstrable antibody response on at least one occasion. The Cp23 ELISA correctly identified 93% of the samples that had a 27-kDa response by Western blot and 100% of the negative samples. The Triton antigen ELISA detected 77% of the samples that had a 17-kDa response by Western blot and 88% of the negative samples. The sensitivity of the Triton antigen assay was higher for samples collected between 16 and 92 days after the onset of symptoms (96%). The minigel-format Western blot did not compare favorably with the large-format blot for the detection of antibodies to the 27-kDa antigen (71% sensitivity). A half-life of about 12 weeks was estimated for antibodies to both the 27- and 17-kDa antigens. We believe the Cp23 and Triton antigen ELISAs will be useful in epidemiologic studies of the prevalence of Cryptosporidium infection in the population.

摘要

微小隐孢子虫是一种原生动物寄生虫,可在包括人类在内的多种哺乳动物宿主中引起腹泻疾病。已证明感染后会产生针对27 kDa和17 kDa大小范围内抗原的特征性血清免疫球蛋白G(IgG)抗体反应,并且已使用几种酶联免疫吸附测定(ELISA)和蛋白质印迹测定方法来测量人血清中的这些IgG水平。我们使用来自实验室确诊的隐孢子虫病患者的一系列样本,比较了两种新ELISA方法与两种不同蛋白质印迹测定方法所获得的结果。当用大型蛋白质印迹法检测时,67名患者中有97%至少在一次检测中呈现出可检测到的抗体反应。Cp23 ELISA正确识别出通过蛋白质印迹法检测到有27 kDa反应的样本中的93%以及所有阴性样本。Triton抗原ELISA检测到通过蛋白质印迹法检测到有17 kDa反应的样本中的77%以及阴性样本中的88%。对于症状出现后16至92天采集的样本,Triton抗原测定的灵敏度更高(96%)。在检测针对27 kDa抗原的抗体方面,小型凝胶蛋白质印迹法与大型蛋白质印迹法相比效果不佳(灵敏度为71%)。估计针对27 kDa和17 kDa抗原的抗体半衰期约为12周。我们认为Cp23和Triton抗原ELISA将有助于人群中隐孢子虫感染流行情况的流行病学研究。

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Cloning of the immunodominant 17-kDa antigen from Cryptosporidium parvum.微小隐孢子虫免疫显性17-kDa抗原的克隆
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