Mary K Athisaya, Hoti S L, Paily K P
Vector Control Research Centre, Indira Nagar, Pondicherry 605 006, India.
J Immunoassay Immunochem. 2007;28(4):343-57. doi: 10.1080/15321810701603484.
Monoclonal antibodies (Mabs) against excretory/secretary (e/s) antigens of fourth stage (L4) larvae of Wuchereria bancrofti were raised and screened for their specificity and sensitivity and evaluated for their potential in detecting homologous e/s antigens in human blood samples. Five Mabs were obtained and, among them, Mab A7 showed high reactivity against e/s antigens of L4 and crude somatic antigens of microfilariae (mf) of W. bancrofti, and infective stage (L3) and adult stage larvae of Brugia malayi. It reacted strongly with sera of Mastomys coucha harbouring L4 stage of B. malayi moderately against sera of the animal having later stages of the parasite. But, it exhibited a low and negligible reactivity against the crude antigens of Setaria cervi and Ascaris lumbricoides, respectively. Another Mab, A6, showed very high reactivity against mf antigens of W. bancrofti and B. malayi and a moderate reactivity against antigens of S. cervi and A. lumbricoides. The two Mabs were tested for their reactivity against filarial antigens in human sera, whose microfilaraemic status was determined by membrane filtration of 1 mL blood sample collected during night. When Mab A7 was tested, 7 out of 22 serum samples (32.0%) from amicrofilaraemic normal individuals from filariasis endemic areas showed positive reactions for filarial antigens, indicating the presence of early stage (L4) of the parasite in them. It also reacted with 84% (n=19) mf positive samples and 11% of non endemic normal serum samples (n=17). Mab A6 showed high reactivity with 86% (n=26) of mf positive serum samples, but did not react with non-endemic normal serum samples (n=17). The results, thus, indicate that the Mab A7 has potential in the detection of e/s antigens of L4 stage larvae of filarial parasites in humans, enabling early diagnosis of filariasis. Mab A6 could be used in the diagnosis of patent infection with microfilaraemia. Western blotting with Mab A7 reacted with the 29.0 kDa protein band of L4 e/s antigens of W. bancrofti.
制备了针对班氏吴策线虫第四期（L4）幼虫排泄/分泌（e/s）抗原的单克隆抗体（Mab），对其特异性和敏感性进行了筛选，并评估了它们在检测人血样本中同源e/s抗原的潜力。获得了5种单克隆抗体，其中，Mab A7对班氏吴策线虫L4的e/s抗原、微丝蚴（mf）的粗体抗原以及马来布鲁线虫感染期（L3）和成虫期幼虫表现出高反应性。它与携带马来布鲁线虫L4期的多乳鼠血清强烈反应，与携带该寄生虫后期的动物血清反应中等。但是，它分别对丝状线虫和蛔虫的粗抗原表现出低反应性且可忽略不计。另一种单克隆抗体A6对班氏吴策线虫和马来布鲁线虫的mf抗原表现出非常高的反应性，对丝状线虫和蛔虫的抗原表现出中等反应性。测试了这两种单克隆抗体对人血清中丝虫抗原的反应性，通过对夜间采集的1 mL血样进行膜过滤来确定其微丝蚴血症状态。当测试Mab A7时，来自丝虫病流行地区无微丝蚴血症正常个体的22份血清样本中有7份（32.0%）对丝虫抗原呈阳性反应，表明其中存在寄生虫的早期阶段（L4）。它还与84%（n = 19）的mf阳性样本和11%的非流行正常血清样本（n = 17）反应。Mab A6与86%（n = 26）的mf阳性血清样本表现出高反应性，但与非流行正常血清样本（n = 17）无反应。因此，结果表明Mab A7在检测人体中丝虫寄生虫L4期幼虫的e/s抗原方面具有潜力，能够实现丝虫病的早期诊断。Mab A6可用于诊断有微丝蚴血症的显性感染。用Mab A7进行的蛋白质印迹法与班氏吴策线虫L4 e/s抗原的29.0 kDa蛋白条带发生反应。
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