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婴儿存档尸检组织中呼吸道合胞病毒核酸的检测

Detection of respiratory syncytial virus nucleic acid in archival postmortem tissue from infants.

作者信息

Cubie H A, Duncan L A, Marshall L A, Smith N M

机构信息

Regional Virus Laboratory, City Hospital, Edinburgh, United Kingdom.

出版信息

Pediatr Pathol Lab Med. 1997 Nov-Dec;17(6):927-38.

PMID:9353832
Abstract

Archival lung tissue from 99 cases of sudden infant death syndrome (SIDS) and from 58 matched comparison cases with known causes of death was studied. Sections were examined by in situ hybridization (ISH) using a cocktail of three synthetic oligonucleotides with sequences chosen from the published sequence of the nucleoprotein gene of respiratory syncytial virus (RS virus). The oligonucleotides were end-labelled with dinitrophenyl (DNP) or digoxigenin (DIG) and hybrids were detected immunocytochemically. RS virus nucleic acid was detected in 24 cases of SIDS (24%) and in 11 (19%) of the comparison group. Specificity was confirmed using a DIG-labeled cloned probe covering the whole of the nucleoprotein gene sequence. With one exception, the same results were obtained. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to confirm the specificity of these results. When matched for age and month and year of death, 76 SIDS cases and 38 controls could be compared. Twenty-one SIDS cases (27.6%) and seven comparison cases (18.4%) contained detectable RS virus sequences by ISH, with a higher detection rate in winter in both groups. The differences were not significant and reflected the seasonal pattern of RS virus infection in the community rather than a causal relationship of RS virus with SIDS. Detection of RS viral mRNA through the summer months suggests that persistence is possible.

摘要

对99例婴儿猝死综合征(SIDS)的存档肺组织以及58例已知死因的匹配对照病例的肺组织进行了研究。使用从呼吸道合胞病毒(RS病毒)核蛋白基因已发表序列中选取的三种合成寡核苷酸混合物,通过原位杂交(ISH)对切片进行检查。这些寡核苷酸用二硝基苯基(DNP)或地高辛配基(DIG)进行末端标记,并通过免疫细胞化学检测杂交体。在24例SIDS病例(24%)和11例(19%)对照组中检测到了RS病毒核酸。使用覆盖整个核蛋白基因序列的DIG标记克隆探针确认了特异性。除一个例外,获得了相同的结果。采用逆转录聚合酶链反应(RT-PCR)来确认这些结果的特异性。当按年龄以及死亡月份和年份进行匹配时,可对76例SIDS病例和38例对照进行比较。通过ISH检测,21例SIDS病例(27.6%)和7例对照病例(18.4%)含有可检测到的RS病毒序列,两组在冬季的检测率均较高。差异不显著,反映的是社区中RS病毒感染的季节性模式,而非RS病毒与SIDS之间的因果关系。在夏季检测到RS病毒mRNA表明其可能持续存在。

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