Kittigul L, Ratanabanangkoon K
Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand.
J Immunoassay. 1993 Sep;14(3):105-27. doi: 10.1080/15321819308019844.
Reverse passive hemagglutination (RPHA) tests for the detection of six major poisonous snake venoms of Thailand were studied. Three different species of red blood cells i.e., sheep (SRBC), human (HRBC) and chicken (CRBC) were sensitized with protein A-affinity purified rabbit antivenom IgG using chromic chloride as a coupling reagent. The properties of these sensitized erythrocytes with regard to sensitivity, specificity, stability to venom enzymes and storage etc., were studied and compared. The sensitivities of the RPHA tests in venom detection were 2 to 635 ng/ml. Cross-reactions were observed with heterologous venoms at concentrations at least 62 times higher than those observed with homologous venoms. After treatment with glutaraldehyde, the coupled red blood cells showed reduced sensitivity but were stable at 4 degrees C from 1 to 12 months depending upon the antibody and the species of erythrocytes. The entire test required 60 to 120 min. The RPHA using fresh SRBC correctly identified various venoms in 48 of 59 (81.3%) serum samples and 16 of 26 (61.5%) wound swabs. Venom mis-identifications were made in 2 sera (3.4%). In a comparison of 24 paired serum and wound swab samples, more positive identifications were made with serum than with swab samples but the difference was not statistically significant (p > 0.05).
对用于检测泰国六种主要毒蛇毒液的反向被动血凝试验（RPHA）进行了研究。使用氯化铬作为偶联剂，用蛋白A亲和纯化的兔抗蛇毒IgG致敏三种不同种类的红细胞，即绵羊红细胞（SRBC）、人红细胞（HRBC）和鸡红细胞（CRBC）。研究并比较了这些致敏红细胞在敏感性、特异性、对毒液酶的稳定性以及储存等方面的特性。RPHA检测毒液的灵敏度为2至635纳克/毫升。观察到与异源毒液有交叉反应，其浓度至少比同源毒液高62倍。用戊二醛处理后，偶联的红细胞敏感性降低，但在4℃下根据抗体和红细胞种类可稳定保存1至12个月。整个试验需要60至120分钟。使用新鲜SRBC的RPHA在59份血清样本中的48份（81.3%）和26份伤口拭子样本中的16份（61.5%）中正确鉴定出了各种毒液。2份血清（3.4%）出现毒液误判。在对24对血清和伤口拭子样本的比较中，血清样本的阳性鉴定结果比伤口拭子样本多，但差异无统计学意义（p>0.05）。
