[Standardization of ELISA IgM and IgA for immunodiagnosis of human trichinosis].

Boletin chileno de parasitologia Pub Date : 1999-07-01
M C Contreras, E Acevedo, S Aguilera, L Sandoval, P Salinas
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Abstract

An ELISA test for trichinosis using as antigen a larvae soluble fraction from Trichinella spiralis was carried out for the detection of IgM and IgA specific antibodies in 45 serum samples from patients confirmed or suspected to have trichinosis by strong clinical and epidemiological evidences. All the patients had positive serology detected by precipitin test, bentonite floculation test, indirect hemagglutination test and ELISA IgG test. The cut-off value was determined using two criteria. Criterion A was determined in each plate, using three positive controls and two negative ones; the average of the negative controls and the weakest positive control, multiplied by a 1.2 factor was, considered the cut-off value. Criterion B was determined using the average plus three standard deviations from 64 apparently healthy persons serum samples. In both cases, three serum dilutions (1:10, 1:100 and 1:500) were used. The sensitivity of ELISA IgM was 100.0, 93.3 and 82.2% using serum dilutions of 1:10, 1:100 and 1:500 respectively (criterion A) and 100.0, 97.8 and 95.6% for the same dilutions (criterion B), whereas the values for ELISA IgA were: 100.0, 91.1 and 86.7% (criterion A) and 100.0, 100.0 and 91.1% (criterion B). In order to find out the specificity of ELISA IgM and ELISA IgA, additional 118 serum samples from individuals with other parasitoses, such as cysticercosis (18) hydatidosis (39), fascioliasis (12), toxocariasis (30), Chagas' disease (12) and individuals with non-specific eosinophilia (7), were also tested. ELISA IgM presented a specificity of 92.3, 93.4 and 97.3% (criterion A) and 96.2, 97.8 and 97.8% (criterion B) whereas the results for ELISA IgA were 97.8, 98.9 and 99.4% (criterion A) and 98.4% for the 1:10 and 1:100 dilutions and 100.0% for the 1:500 dilution (criterion B). The positive predictive values of ELISA IgM were 76.3, 77.8 and 88.1% (criterion A) and 86.5, 91.7 and 91.5% (criterion B) whereas the negative ones were 100.0, 98.3 and 95.7% (criterion A) and 100.0, 99.4 and 98.9% (criterion B). The positive predictive values of ELISA IgA were 91.8, 95.3 and 97.5% (criterion A) and 93.8, 93.8 and 100.0% (criterion B) whereas the negatives ones were: 100.0, 97.8 and 96.8% (criterion A) and 100.0, 100.0 and 97.8% (criterion B). The use of ELISA IgM and ELISA IgA in the immunodiagnosis of trichinosis is discussed.

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[人旋毛虫病ELISA IgM和IgA免疫诊断的标准化]。
采用旋毛虫幼虫可溶性部分为抗原的旋毛虫病酶联免疫吸附试验(ELISA),对45份有临床和流行病学证据的确诊或疑似旋毛虫病患者血清样本进行IgM和IgA特异性抗体检测。沉淀试验、膨润土絮凝试验、间接血凝试验、ELISA IgG检测均为血清阳性。临界值采用两个标准确定。每个平板测定标准A,采用3个阳性对照和2个阴性对照;阴性对照和最弱阳性对照的平均值乘以一个1.2因子,被认为是临界值。标准B是用64个明显健康的人血清样本的平均值加3个标准差确定的。在这两种情况下,使用三种血清稀释(1:10,1:100和1:500)。ELISA IgM在血清稀释度为1:10、1:100和1:500时的敏感性分别为100.0、93.3和82.2%(标准A),在相同稀释度下的敏感性分别为100.0、97.8和95.6%(标准B),而ELISA IgA的敏感性分别为:100.0、91.1和86.7%(标准A)和100.0、100.0和91.1%(标准B)。为了确定ELISA IgM和ELISA IgA的特异性,我们还检测了其他寄生虫病患者的血清样本118份,如囊虫病(18份)、包虫病(39份)、片吸虫病(12份)、弓形虫病(30份)、恰加斯病(12份)和非特异性嗜酸粒细胞增多症(7份)。ELISA IgM特异性为92.3,93.4和97.3%(标准)和96.2,97.8和97.8%(标准B)而ELISA IgA的结果是97.8,98.9和99.4%(标准),98.4%为1:10和1:10 0稀释和100.0% 1:50 0稀释(标准B)。ELISA IgM阳性预测值的76.3,77.8和88.1%(标准)和86.5,91.7和91.5%(标准B)而消极的是100.0,98.3和95.7%(标准)和100.0,ELISA IgA阳性预测值分别为91.8、95.3、97.5%(判据A)和93.8、93.8、100.0%(判据B),阴性预测值分别为100.0、97.8、96.8%(判据A)和100.0、100.0、97.8%(判据B)。探讨ELISA IgM和ELISA IgA在旋毛虫病免疫诊断中的应用。
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