Department of Virology II, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
J Clin Microbiol. 2010 Aug;48(8):2698-702. doi: 10.1128/JCM.00207-10. Epub 2010 Jun 2.
In the Global Polio Eradication Initiative, laboratory diagnosis plays a critical role by isolating and identifying poliovirus (PV) from the stool samples of patients with acute flaccid paralysis (AFP). In this study, we developed a particle agglutination (PA) method with a soluble human PV receptor (hPVR) in the form of an immunoadhesin (PVR-IgG2a) for the simple and rapid identification of PV. Sensitized gelatin particles with PVR-IgG2a showed specific agglutination with the culture fluid of PV-infected cells within 2 h of reaction in a one-step procedure. Detection limits for type 1, 2, and 3 PV(Sabin) strains were 1.5 x 10(6) 50% cell culture infectious doses (CCID(50)), 5.3 x 10(5) CCID(50), and 9.1 x 10(5) CCID(50), respectively. Wild-type PVs and PV isolates from acute flaccid paralysis cases examined were identified correctly with this PA method, except for some samples with a mixture of different serotypes of PVs, where a minor population of PV failed to be detected. These results suggest that this PA method is useful for the simple and rapid identification of PV, although the sensitivity was not high enough to detect a minor population of PV (<1/10 of the major population) among mixed PVs.
在全球消灭脊灰炎行动中,实验室诊断通过从急性弛缓性麻痹(AFP)患者的粪便样本中分离和鉴定脊灰病毒(PV)发挥着关键作用。在这项研究中,我们开发了一种粒子聚集(PA)方法,该方法使用可溶性人 PV 受体(hPVR)作为免疫黏附素(PVR-IgG2a),用于简单快速地鉴定 PV。用 PVR-IgG2a 敏化的明胶颗粒在一步反应中,与感染细胞的培养液在 2 小时内表现出特异性聚集。1 型、2 型和 3 型 PV(Sabin)株的检测限分别为 1.5 x 10(6)50%细胞培养感染剂量(CCID(50))、5.3 x 10(5)CCID(50)和 9.1 x 10(5)CCID(50)。除了一些含有不同血清型 PV 混合物的样本外,这种 PA 方法能够正确识别野生型 PV 和急性弛缓性麻痹病例中分离的 PV,在这些样本中,PV 的一个小群体未能被检测到。这些结果表明,尽管该 PA 方法的敏感性不足以检测到混合 PV 中(主要群体的 1/10 以下)的 PV 小群体,但它对于简单快速地鉴定 PV 非常有用。