Department of Veterinary Parasitology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, 37, Kshudiram Bose Sarani, Kolkata 700 037, India.
Vet Parasitol. 2010 Jun 24;170(3-4):262-7. doi: 10.1016/j.vetpar.2010.02.013. Epub 2010 Feb 18.
A polyclonal antibody based coproantigen detection enzyme linked immunosorbent assay (cAg-ELISA) for diagnosis of experimental and natural Oesophagostomum columbianum infection in goats was developed and evaluated. Adult O. columbianum worms, collected from the caecum and colon of slaughtered goats, were triturated and cultured for obtaining infective third stage larvae (L(3)) and also used for preparation of excretory-secretory antigen (ESAg). Experimental goats were orally infected each with 600 L(3)/kg of the body weight. Filter sterilized faecal supernatant, i.e. the coproantigen (cAg) was harvested from the rectal faeces of all the infected goats, on alternate days from day-5 till day-31 after the infection. Hyperimmune serum (HIS) against ESAg of O. columbianum was raised in rabbits. Molecular and antigenic characterization of ES products of O. columbianum by HIS revealed that 50 and 39kDa polypeptides were immuno-dominant. Coproantigen detection ELISA was standardized by using the cAg as coating antigen and its subsequent binding with the HIS against ESAg of O. columbianum. The sensitivity, specificity and accuracy of the standardized assay were determined by evaluating the assay on the faecal supernatant of 96 slaughtered goats taking into consideration their recorded parasitological status in respect of the abomasal and the intestinal parasites. The cAg-ELISA detected the prepatent oesophagostomosis on 20-24-day-post-infection with a sensitivity, specificity and accuracy of 88, 89.13 and 88.54%, respectively. The assay is relatively easy to perform and would serve as a reliable tool for detection of caprine nodular oesophagostomosis.
一种基于多克隆抗体的共抗原检测酶联免疫吸附试验(cAg-ELISA)被开发并评估用于诊断山羊实验和自然感染食道口线虫。从屠宰山羊的盲肠和结肠中收集的成年食道口线虫蠕虫被研磨并培养以获得感染性第三期幼虫(L(3)),并用于制备排泄分泌抗原(ESAg)。实验山羊每只口服感染 600 条 L(3)/kg 体重。从感染后第 5 天到第 31 天,从所有感染的山羊的直肠粪便中采集过滤消毒的粪便上清液,即共抗原(cAg)。针对食道口线虫 ESAg 的高免血清(HIS)在兔子中产生。HIS 对食道口线虫 ES 产物的分子和抗原特性分析表明,50 和 39kDa 多肽是免疫显性的。使用 cAg 作为包被抗原并随后与针对食道口线虫 ESAg 的 HIS 结合,对共抗原检测 ELISA 进行了标准化。通过评估考虑到屠宰山羊胃和肠道寄生虫记录的寄生虫学状况的 96 份粪便上清液,确定了标准化测定的敏感性、特异性和准确性。cAg-ELISA 在感染后 20-24 天检测到未成熟的食道口线虫病,敏感性、特异性和准确性分别为 88%、89.13%和 88.54%。该测定相对容易进行,将成为检测山羊结节性食道口线虫病的可靠工具。