McLauchlin J, Casemore D P, Moran S, Patel S
Food Hygiene Laboratory, PHLS Central Public Health Laboratory, London, UK.
Folia Parasitol (Praha). 1998;45(2):83-92.
A technique based on the analysis of banding patterns obtained by SDS-PAGE Western-blotting of an oocyst wall antigen obtained from faeces has been evaluated to subtype Cryptosporidium parvum Tyzzer, 1912. This technique appears to have sufficient stability to recognise multiple types of this parasite. A similar Western-blotting technique has also been used to assess antibody responses to cryptosporidial antigens in human sera. Two systems were developed: one against three antigens of apparent molecular weights 6, 14 and 17 kDa; the second against oocyst wall antigens of apparent molecular weights 57, 69, 75, 89, 128, 151 and 173 kDa. Antibodies to three antigens of apparent molecular weights 6, 14 and 17 kDa were most successful as diagnostic markers in that they were found in > 88% of convalescent phase sera from confirmed cryptosporidiosis patients and were uncommon (> or = 7%) in control subjects. Faecal samples from human and animal sporadic cases yielded a wide range of cryptosporidial antigen banding patterns. Samples from patients in a water-borne outbreak in South Devon (England) in 1995 also yielded a wide range of banding patterns including members within individual household family groups. These results are in contrast with those from samples collected from other defined geographical areas, including some from a second water-borne outbreak where much more homogeneous banding patterns were obtained. Sera collected for other purposes from apparently uninfected individuals 9 months after the South Devon 1995 outbreak were examined. Antibodies to the three antigens of molecular weights 6, 14 and 17 kDa were detected in 32-49% of individuals resident in the outbreak water supply area, and in 15-21% of those resident in an adjacent water supply area. The significance of these findings is discussed in relation to data obtained from epidemiological field studies.
一种基于对从粪便中获得的卵囊壁抗原进行SDS - PAGE免疫印迹分析所得到的条带模式的技术，已被用于对微小隐孢子虫（Tyzzer，1912）进行亚型分析。该技术似乎具有足够的稳定性来识别这种寄生虫的多种类型。一种类似的免疫印迹技术也已被用于评估人血清中针对隐孢子虫抗原的抗体反应。开发了两种系统：一种针对表观分子量为6、14和17 kDa的三种抗原；第二种针对表观分子量为57、69、75、89、128、151和173 kDa的卵囊壁抗原。针对表观分子量为6、14和17 kDa的三种抗原的抗体作为诊断标志物最为成功，因为它们在确诊的隐孢子虫病患者恢复期血清中的检出率> 88%，而在对照受试者中不常见（>或 = 7%）。来自人类和动物散发病例的粪便样本产生了广泛的隐孢子虫抗原条带模式。1995年在英国南德文郡一次水源性暴发中患者的样本也产生了广泛的条带模式，包括各个家庭群体中的成员。这些结果与从其他特定地理区域收集的样本结果形成对比，包括一些来自第二次水源性暴发的样本，在那里获得了更为一致的条带模式。对1995年南德文郡暴发9个月后从明显未感染个体收集的用于其他目的的血清进行了检测。在暴发供水区域居住的个体中，32 - 49%检测到针对分子量为6、14和17 kDa的三种抗原的抗体，在相邻供水区域居住的个体中，这一比例为15 - 21%。结合从流行病学现场研究获得的数据对这些发现的意义进行了讨论。
