Xiao L, Herd R P
Department of Veterinary Preventive Medicine, Ohio State University College of Veterinary Medicine, Columbus 43210.
J Clin Microbiol. 1993 Nov;31(11):2944-6. doi: 10.1128/jcm.31.11.2944-2946.1993.
The lack of quick, simple, and sensitive quantitative tests has impeded studies on infection patterns and treatment of Giardia spp. and Cryptosporidium spp. A quantitative direct immunofluorescence assay (FA) using a commercial FA kit was developed and evaluated. Recovery rates of the FA for Cryptosporidium oocysts in calf feces seeded with 1,000, 10,000, 100,000, and 1,000,000 oocysts per g were 14.8, 40.8, 84.2, and 78.2%, respectively. Interassay coefficients of variation were 10.6 to 47.1%. Recovery rates of the FA for Giardia cysts in feces seeded with 1,000, 10,000, and 100,000 cysts per g were 76.4, 96.9, and 89.6%, respectively. Interassay coefficients of variation were 7.4 to 22.1%. By comparison, recovery rates of Giardia cyst by sucrose gradient flotation were only 20.5, 51.2, and 42.9%, respectively. Counts of cysts-per-gram obtained by sucrose gradient flotation with samples from calves, lambs, and ewes were only 49.1 to 54.8% of those obtained by the FA. Zinc sulfate flotation detected only 36.4% of infections when there were < or = 1,000 cysts per g. The quantitative FA offers a useful technique for epidemiological and control studies of these two parasites.
缺乏快速、简便且灵敏的定量检测方法阻碍了对贾第虫属和隐孢子虫属感染模式及治疗的研究。我们开发并评估了一种使用商用直接免疫荧光检测试剂盒的定量直接免疫荧光检测法(FA)。在每克接种1000、10000、100000和1000000个卵囊的小牛粪便中,隐孢子虫卵囊的FA回收率分别为14.8%、40.8%、84.2%和78.2%。批间变异系数为10.6%至47.1%。在每克接种1000、10000和100000个包囊的粪便中,贾第虫包囊的FA回收率分别为76.4%、96.9%和89.6%。批间变异系数为7.4%至22.1%。相比之下,通过蔗糖梯度浮选法检测贾第虫包囊的回收率分别仅为20.5%、51.2%和42.9%。用来自小牛、羔羊和母羊的样本通过蔗糖梯度浮选法获得的每克包囊计数仅为FA法获得计数的49.1%至54.8%。当每克包囊数≤1000个时,硫酸锌浮选法仅能检测到36.4%的感染情况。这种定量FA法为这两种寄生虫的流行病学和防治研究提供了一种有用的技术。