用于血清学诊断西部马脑炎病毒所致人类感染的斑点酶联免疫吸附测定。

Dot-ELISA for serodiagnosis of human infections due to Western equine encephalitis virus.

作者信息

Boctor F N, Calisher C H, Peter J B

机构信息

Specialty Laboratories, Inc., Santa Monica, CA 90404-3900.

出版信息

J Virol Methods. 1989 Dec;26(3):305-11. doi: 10.1016/0166-0934(89)90112-2.

DOI:10.1016/0166-0934(89)90112-2

PMID:2621241

Abstract

A standard dot-ELISA (enzyme-linked immunosorbent assay) was modified for use in detecting IgM and IgG class antibodies to Western equine encephalitis (WEE) virus in serum samples from humans infected with this virus. Nitrocellulose membranes were soaked in supernatant fluid from WEE virus-infected cell cultures, air dried, and blocked with bovine protein. Serum samples were pipetted onto sections of the nitrocellulose, incubated, and washed. Addition of antibody to human immunoglobulin conjugated to alkaline phosphatase and enzyme substrate were used to detect the antibodies. Of 13 samples positive for IgM antibody to WEE virus by IgM antibody capture ELISA, 12 were positive by IgM dot-ELISA. IgG antibody to WEE virus was detected by dot-ELISA in 7/8, 10/14 and 7/10 samples with neutralizing, hemagglutination-inhibiting, or complement-fixing antibodies, respectively.

摘要

一种标准的斑点酶联免疫吸附测定（dot-ELISA）被改良用于检测感染西部马脑炎（WEE）病毒的人类血清样本中针对该病毒的IgM和IgG类抗体。将硝酸纤维素膜浸泡在感染WEE病毒的细胞培养物的上清液中，空气干燥，并用牛血清白蛋白封闭。将血清样本移液到硝酸纤维素膜切片上，孵育并洗涤。加入与碱性磷酸酶偶联的抗人免疫球蛋白抗体和酶底物来检测抗体。在通过IgM抗体捕获ELISA检测为WEE病毒IgM抗体阳性的13份样本中，有12份通过IgM斑点酶联免疫吸附测定呈阳性。分别在具有中和抗体、血凝抑制抗体或补体结合抗体的7/8、10/14和7/10份样本中，通过斑点酶联免疫吸附测定检测到了针对WEE病毒的IgG抗体。