Mathiesen L R, Feinstone S M, Wong D C, Skinhoej P, Purcell R H
J Clin Microbiol. 1978 Feb;7(2):184-93. doi: 10.1128/jcm.7.2.184-193.1978.
Previously described techniques for detection of hepatitis A antigen (HA Ag) and antibody (anti-HA) have required purified HA Ag and expensive equipment. Herein is described an enzyme-linked immunosorbent assay (ELISA) for specific detection of HA Ag in human stool filtrates and of anti-HA in sera by using selected HA Ag-containing human stool filtrates as the antigen source. Because human stools often react nonspecifically in serological tests for HA Ag, blocking with preexposure and hyperimmune anti-HA sera from a chimpanzee inoculated with hepatitis A virus was used to confirm specific detection of HA Ag. The sensitivity of ELISA was found to be comparable to that of solid-phase radioimmunoassay (SPRIA) and immune electron microscopy (IEM). Of 37 acute-phase stools collected from nine patients, 16 were positive for HA Ag by ELISA. In 13 of these, HA Ag particles were found by IEM, and an additional 3 stools negative by ELISA contained HA Ag particles by IEM. Eight control stools were negative by both ELISA and IEM. Anti-HA was measured in sera by demonstrating its ability to block binding of the enzyme conjugate to HA Ag in a stool without detectable nonspecificity. This test (blocking ELISA) was as sensitive and specific as blocking SPIRA, IEM, and immune adherence hemagglutination and, like SPRIA and IEM, detected early-developing antibody. The ELISA is simple to perform and requires only a minimum of equipment. It is useful for screening stools for HA Ag and for monitoring HA Ag during purification, as well as for detecting early and late anti-HA in sera.
先前描述的用于检测甲型肝炎抗原（HA Ag）和抗体（抗-HA）的技术需要纯化的HA Ag和昂贵的设备。本文描述了一种酶联免疫吸附测定（ELISA），通过使用选定的含HA Ag的人粪便滤液作为抗原来源，特异性检测人粪便滤液中的HA Ag和血清中的抗-HA。由于人粪便在HA Ag的血清学检测中常出现非特异性反应，因此使用来自接种甲型肝炎病毒的黑猩猩的预免疫和超免疫抗-HA血清进行封闭，以确认HA Ag的特异性检测。发现ELISA的灵敏度与固相放射免疫测定（SPRIA）和免疫电子显微镜（IEM）相当。从9名患者收集的37份急性期粪便中，有16份通过ELISA检测HA Ag呈阳性。其中13份通过IEM发现了HA Ag颗粒，另外3份ELISA检测为阴性的粪便通过IEM含有HA Ag颗粒。8份对照粪便通过ELISA和IEM检测均为阴性。通过证明血清中抗-HA能够在无明显非特异性的情况下阻断酶结合物与粪便中HA Ag的结合来测量抗-HA。该试验（阻断ELISA）与阻断SPIRA、IEM和免疫粘附血凝试验一样灵敏和特异，并且与SPRIA和IEM一样，能检测到早期产生的抗体。ELISA操作简单，只需要最少的设备。它可用于筛查粪便中的HA Ag，在纯化过程中监测HA Ag，以及检测血清中早期和晚期的抗-HA。
