Livingston R S, Riley L K, Steffen E K, Besch-Williford C L, Hook R R, Franklin C L
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia 65211, USA.
J Clin Microbiol. 1997 May;35(5):1236-8. doi: 10.1128/jcm.35.5.1236-1238.1997.
Helicobacter hepaticus is a newly recognized bacterium associated with chronic active hepatitis, hepatic carcinoma, and inflammatory bowel disease in mice. Currently, fecal or tissue PCR, fecal culture, or histologic examination of silver-stained liver sections is used to diagnose H. hepaticus infection. In this report, we describe an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of H. hepaticus infection in mice with a membrane digest preparation of H. hepaticus as the antigen. Sera from mice positive for H. hepaticus by PCR or histologic examination (n = 88), positive for Helicobacter bilis by PCR (n = 13), positive for other helicobacters (not identifiable to species level) by PCR (n = 25), or negative for all Helicobacter species by PCR (n = 162) were used to evaluate the ELISA. Results indicated that ELISA provided 93.2% sensitivity, 94% specificity, 87.2% positive predictive value, and 96.9% negative predictive value. Cross-reactive antibodies were detected in some mice infected with helicobacters not identifiable to species level. To further define ELISA sensitivity and specificity, groups of 10 C57BL/6 mice were inoculated per os with H. hepaticus, Helicobacter muridarum, or H. bilis. Sera were collected and examined by the ELISA. H. hepaticus-infected mice seroconverted by 2 weeks and maintained ELISA reactivity throughout the 18-week study, while mice infected with H. muridarum and H. bilis were negative by ELISA. These results indicate that this reported ELISA is highly sensitive and specific for the serodiagnosis of H. hepaticus infection in mice.
肝螺杆菌是一种新发现的细菌,与小鼠的慢性活动性肝炎、肝癌和炎症性肠病有关。目前,粪便或组织聚合酶链反应(PCR)、粪便培养或银染肝切片的组织学检查用于诊断肝螺杆菌感染。在本报告中,我们描述了一种酶联免疫吸附测定(ELISA)方法,以肝螺杆菌的膜消化制剂为抗原,用于小鼠肝螺杆菌感染的血清学诊断。使用通过PCR或组织学检查确诊为肝螺杆菌阳性的小鼠血清(n = 88)、通过PCR确诊为胆汁螺杆菌阳性的小鼠血清(n = 13)、通过PCR确诊为其他螺杆菌(无法鉴定到种水平)阳性的小鼠血清(n = 25)或通过PCR确诊所有螺杆菌属均为阴性的小鼠血清(n = 162)来评估该ELISA方法。结果表明,ELISA的灵敏度为93.2%,特异性为94%,阳性预测值为87.2%,阴性预测值为96.9%。在一些感染了无法鉴定到种水平的螺杆菌的小鼠中检测到了交叉反应抗体。为了进一步确定ELISA的灵敏度和特异性,每组10只C57BL/6小鼠经口接种肝螺杆菌、鼠螺杆菌或胆汁螺杆菌。收集血清并通过ELISA进行检测。感染肝螺杆菌的小鼠在2周时血清转化,并在整个18周的研究中保持ELISA反应性,而感染鼠螺杆菌和胆汁螺杆菌的小鼠ELISA检测为阴性。这些结果表明,本报告中的ELISA方法对小鼠肝螺杆菌感染的血清学诊断具有高度的敏感性和特异性。