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[一所智障机构中甲型肝炎病毒引起的暴发——采用十六烷基三甲基溴化铵法检测甲型肝炎病毒核糖核酸]

[An outbreak caused by hepatitis A virus in an institution for the mentally handicapped--detection of hepatitis A virus RNA using CTAB method].

作者信息

Kitahashi T, Tanaka T, Ishikawa Y, Hasegawa S

机构信息

Chiba City Institute of Health and Environment.

出版信息

Kansenshogaku Zasshi. 1998 Aug;72(8):794-800. doi: 10.11150/kansenshogakuzasshi1970.72.794.

Abstract

The prevalence of hepatitis A virus (HAV) antibody in people has decreased from year to year in Japan. A sequential outbreak occurred in an institution for the mentally handicapped people in Chiba City in the summer of 1995. Eight people were infected including 7 residents and one staff member. We tested to detect antigen in fecal samples by ELISA and PCR for early diagnosis for hepatitis A infection. Four sera and 5 feces were obtained from 5 patients between 2 and 8 days after the onset of symptoms. The anti-HAV IgM was found to be positive in 4 sera examined. The HAV antigen was detected in 3 out of 5 feces using ELISA. An existence of inhibitor in 2 negative specimens against the ELISA was suggested by the recovery test of added antigen. HAV RNA was extracted by CTAB method from feces and detected in 4 our of 5 specimens in PCR amplification and in all of 5 specimens in nested PCR amplification. The sequence of PCR products in the P1/P2 junction of the HAV genome revealed that the virus associated with the outbreak belongs to HAV subgenotype IA. HAV RNA was detected in ELISA negative specimens and in the specimen from a patient 2 days after the onset of symptoms using PCR amplification by CTAB method. These results indicate that PCR amplification was useful for the early diagnosis of hepatitis A infection.

摘要

在日本,人群中甲肝病毒(HAV)抗体的流行率逐年下降。1995年夏季,千叶市一所智障人士机构发生了连续性疫情。8人被感染,其中包括7名居民和1名工作人员。为进行甲型肝炎感染的早期诊断,我们通过ELISA和PCR检测粪便样本中的抗原。在症状出现后的2至8天内,从5名患者身上采集了4份血清和5份粪便。在所检测的4份血清中,抗-HAV IgM呈阳性。使用ELISA在5份粪便中的3份中检测到了HAV抗原。通过添加抗原的回收率试验表明,2份ELISA阴性标本中存在抑制剂。采用CTAB法从粪便中提取HAV RNA,在PCR扩增中5份标本中的4份以及巢式PCR扩增中5份标本全部检测到了HAV RNA。HAV基因组P1/P2连接处PCR产物的序列显示,与此次疫情相关的病毒属于HAV IA亚基因型。采用CTAB法通过PCR扩增在ELISA阴性标本以及症状出现后2天患者的标本中检测到了HAV RNA。这些结果表明,PCR扩增对甲型肝炎感染的早期诊断很有用。

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