Strauss W, O'Neill S M, Parkinson M, Angles R, Dalton J P
Unidad de Parasitologie, Instituto Nacional de Laboratorios de Salud, Secretaria Nacional de Salud, La Paz, Bolivia.
Am J Trop Med Hyg. 1999 May;60(5):746-8. doi: 10.4269/ajtmh.1999.60.746.
We have developed an ELISA for the diagnosis of human fascioliasis based on the detection of IgG4 antibodies to Fasciola hepatica cathepsin LI cysteine protease. Use of this assay in the Bolivian Altiplano, a region with a high prevalence of the disease, was hampered by the reluctance of the indigenous population to provide blood. To overcome this problem, we have investigated the method of collecting small quantities of blood from the finger onto filter paper, followed by the elution of antibodies for use in the diagnostic assay. Serum samples and blood samples collected onto filter paper were obtained from 57 individuals living in the village of Cutusuma in 1987 and from 11 individuals in Chijipata in 1996. Analysis of the IgG4-ELISA results revealed that there is highly significant linear relationship (P < 0.001) between the two methods of sampling. Most importantly, a reliable diagnosis was made with the blood-filter samples from Cutusuma, which had been stored for 10 years at 40 degrees C. While some deterioration of the blood-filter samples from Cutusuma had occurred over the 10-year storage period, no deterioration occurred with the Chijipata samples, which were stored for one year. Therefore, the method of collecting blood onto filter paper should prove useful for large-scale epidemiologic studies on human fascioliasis in the Bolivian Altiplano and in other regions where this disease is prevalent.
我们基于检测针对肝片吸虫组织蛋白酶L1半胱氨酸蛋白酶的IgG4抗体,开发了一种用于诊断人体肝片吸虫病的酶联免疫吸附测定(ELISA)。在疾病高发的玻利维亚高原地区使用该检测方法时,当地居民不愿提供血液,这给检测带来了阻碍。为克服这一问题,我们研究了从手指采集少量血液到滤纸上,然后洗脱抗体用于诊断检测的方法。1987年从库图苏马村的57个人以及1996年从奇吉帕塔的11个人那里获取了血清样本和采集到滤纸上的血液样本。对IgG4-ELISA结果的分析表明,两种采样方法之间存在高度显著的线性关系(P < 0.001)。最重要的是,对在40摄氏度下保存了10年的库图苏马血液滤纸样本做出了可靠诊断。虽然库图苏马血液滤纸样本在10年的保存期内出现了一些降解,但保存了一年的奇吉帕塔样本没有发生降解。因此,将血液采集到滤纸上的方法对于玻利维亚高原地区以及其他该疾病流行地区的人体肝片吸虫病大规模流行病学研究应是有用的。