Salminen K K, Vuorinen T, Oikarinen S, Helminen M, Simell S, Knip M, Ilonen J, Simell O, Hyöty H
Juvenile Diabetes Research Foundation Center for Prevention of Type 1 Diabetes in Finland.
Diabet Med. 2004 Feb;21(2):156-64. doi: 10.1111/j.1464-5491.2004.01097.x.
To develop methods for isolation of enterovirus strains from subjects with preclinical Type 1 diabetes and evaluate if their presence in stools is associated with beta-cell damage.
The study subjects were participants of the Finnish Type 1 Diabetes Prediction and Prevention Study (DIPP). The prospectively followed birth cohort comprised 12 children who turned positive for diabetes-associated autoantibodies during the follow-up (case children) and 53 controls matched for date of birth, sex and HLA-DQB1 alleles. Altogether, 878 stool samples were analysed for the presence of enterovirus RNA by RT-PCR followed by virus isolation and partial sequencing of viral genome. Enterovirus antibodies and RNA were simultaneously analysed from serum.
Eleven enterovirus infections were diagnosed in case children and 42 infections in control children by the presence of viral RNA in stools. The proportion of children who were repeatedly enterovirus RNA-positive stools was higher among case than control children (42% vs. 11% of children; P=0.02). Combined serum (antibody and RT-PCR) and stool analyses indicated at least one enterovirus infection in 83% of the case children before the appearance of autoantibodies, while only 42% of the control children had infection by the same age (P=0.006). Twelve enterovirus strains were isolated from case children and 38 strains from control children.
This protocol makes it possible to isolate a large number of enterovirus strains from prediabetic subjects. The findings suggest that enterovirus infections may be associated with the beta-cell damaging process.
开发从临床前期1型糖尿病患者中分离肠道病毒株的方法，并评估其在粪便中的存在是否与β细胞损伤有关。
研究对象为芬兰1型糖尿病预测与预防研究（DIPP）的参与者。前瞻性随访的出生队列包括12名在随访期间糖尿病相关自身抗体转为阳性的儿童（病例儿童）和53名在出生日期、性别和HLA - DQB1等位基因方面匹配的对照儿童。总共对878份粪便样本进行逆转录聚合酶链反应（RT - PCR）分析以检测肠道病毒RNA的存在，随后进行病毒分离及病毒基因组的部分测序。同时从血清中分析肠道病毒抗体和RNA。
通过粪便中病毒RNA的存在，在病例儿童中诊断出11例肠道病毒感染，在对照儿童中诊断出42例感染。病例儿童中粪便多次肠道病毒RNA呈阳性的儿童比例高于对照儿童（分别为42%和11%；P = 0.02）。血清（抗体和RT - PCR）与粪便联合分析表明，83%的病例儿童在自身抗体出现之前至少有一次肠道病毒感染，而在同一年龄时，只有42%的对照儿童有感染（P = 0.006）。从病例儿童中分离出12株肠道病毒株，从对照儿童中分离出38株。
该方案使得从糖尿病前期受试者中分离出大量肠道病毒株成为可能。研究结果表明肠道病毒感染可能与β细胞损伤过程有关。
