Sri Sakthi Amma Institute of Biomedical Research, Sri Narayani Hospital & Research Centre, Vellore, India.
Department of Microbiology, King George Medical University, Lucknow, India.
Indian J Med Res. 2017 Sep;146(3):381-385. doi: 10.4103/ijmr.IJMR_225_16.
BACKGROUND & OBJECTIVES: Parvovirus B19 infections occur worldwide; the infection is acquired early in childhood but could occur later. B19 is reported to cause infection in childhood febrile illnesses, and arthropathies in adults and children and in end-stage renal disease (ESRD) seen in adults. This study was designed to develop an in-house IgM indirect ELISA for serological screening among patients and controls, and to compare ELISA results with those of nested polymerase chain reaction (nPCR) assay.
An in-house IgM indirect ELISA was standardized using peptide sequence of VP1/VP2 region of parvovirus B19. A total of 201 children and adult with febrile illnesses, 216 individuals with non-traumatic arthropathies, 201 cases of chronic anaemia associated with ESRD and 100 healthy controls were tested. Serum was separated from the blood and subsequently used for DNA extraction. The nested polymerase chain reaction (nPCR) for the detection of B19V DNA was performed using primers targeting the overlapping region of VP1/VP2 capsid protein genes.
A total of 618 samples were tested for parvovirus B19 by an in-house IgM indirect ELISA. Among these samples, six were positive by in-house ELISA. The inter-rater agreement between ELISA and PCR assays was calculated using kappa coefficient analysis. The value of κ was 0.77 and the strength of agreement was 'good' (P<0.001).
INTERPRETATION & CONCLUSIONS: The in-house IgM indirect ELISA was found to be simple with high sensitivity and specificity when compared with nPCR and could be used as an alternative to expensive commercial kits in resource-poor settings.
细小病毒 B19 感染遍布全球；感染通常发生在儿童早期，但也可能发生在后期。B19 据报道会导致儿童发热性疾病感染，并导致成人和儿童出现关节病以及成人终末期肾病（ESRD）。本研究旨在开发一种内部 IgM 间接 ELISA 用于患者和对照者的血清学筛查，并将 ELISA 结果与巢式聚合酶链反应（nPCR）检测结果进行比较。
使用细小病毒 B19 的 VP1/VP2 区域的肽序列来标准化内部 IgM 间接 ELISA。共检测了 201 名发热疾病的儿童和成人、216 名非创伤性关节病患者、201 例与 ESRD 相关的慢性贫血病例和 100 名健康对照者。从血液中分离血清，随后用于 DNA 提取。使用针对 VP1/VP2 衣壳蛋白基因重叠区的引物进行巢式聚合酶链反应（nPCR）检测 B19V DNA。
共对 618 个样本进行了内部 IgM 间接 ELISA 检测。其中 6 个样本经内部 ELISA 检测呈阳性。使用 Kappa 系数分析计算 ELISA 和 PCR 检测之间的观察者间一致性。κ 值为 0.77，一致性强度为“良好”（P<0.001）。
与 nPCR 相比，内部 IgM 间接 ELISA 具有较高的灵敏度和特异性，并且简单，可在资源匮乏的环境中替代昂贵的商业试剂盒。
