Verification of the reactivity of immunoglobulins in dried blood spots collected for onchocerciasis sero-surveillance by an Escherichia coli ELISA

H. Hassan, Kristi Miley, T. Unnasch
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Abstract

The World Health Organization guidelines for verification of onchocerciasis elimination include demonstrating that the prevalence of exposure to the parasite in individuals born since transmission was interrupted needs to be less than 0.1%. The guidelines recommend using seropositivity to an Onchocerca volvulus specific antigen (Ov16) for this purpose. Ov16 seropositivity has most often been assessed using the Ov16 ELISA assay. Currently, the Ov16 ELISA assay includes internal positive and negative controls to monitor for proper assay performance but does not control for the quality of the dried blood spots (DBS) being tested. Previous studies have reported a high prevalence of antibodies recognizing Escherichia coli in children. Through the development of an ELISA assay to detect antibodies recognizing E. coli, a common commensal in humans, DBS may be prescreened for quality assurance prior to testing for Ov16. Results demonstrated antibodies to E. coli were detected in 100% of randomly selected serum samples collected from O. volvulus infected individuals residing in an onchocerciasis hyperendemic area. Furthermore, when DBS were improperly stored, the E. coli antibodies were found to decay over a period of one week, while remaining unchanged over the same period in properly stored samples. Similarly, E. coli antibodies were detected in 100% of a batch of field collected properly stored DBS, while being present only in 5% of a batch of improperly stored spots. This study demonstrates the value of E. coli ELISA for DBS quality control testing and validation of proper storage of collections of DBS for the Ov16 ELISA.
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用大肠杆菌酶联免疫吸附试验验证盘尾丝虫病血清监测采集的干血斑中免疫球蛋白的反应性
世界卫生组织的消除盘尾丝虫病验证准则包括:证明自传播中断后出生的个体中接触寄生虫的流行率需低于 0.1%。指导方针建议使用盘尾丝虫特异性抗原(Ov16)血清阳性来达到这一目的。Ov16血清阳性通常使用Ov16 ELISA检测法进行评估。目前,Ov16 酶联免疫吸附测定法包括内部阳性和阴性对照,用于监测测定结果的正确性,但不能控制被测干血斑点(DBS)的质量。先前的研究报告显示,儿童体内识别大肠杆菌的抗体流行率很高。通过开发一种酶联免疫吸附试验(ELISA)来检测大肠杆菌(一种人类常见的共生菌)抗体,可以在检测 Ov16 之前对干血斑进行预筛,以保证质量。结果表明,在盘尾丝虫病高发区随机采集的大肠杆菌感染者血清样本中,100%都能检测到大肠杆菌抗体。此外,当 DBS 储存不当时,大肠杆菌抗体会在一周内衰减,而正确储存的样本在同一时期内则保持不变。同样,在一批现场采集并妥善保存的 DBS 样品中,100% 都能检测到大肠杆菌抗体,而在一批保存不当的样本中,只有 5% 的样本能检测到大肠杆菌抗体。这项研究证明了大肠杆菌酶联免疫吸附试验在 DBS 质量控制检测中的价值,并验证了 Ov16 酶联免疫吸附试验中 DBS 的正确储存。
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