Takami T, Kawashima H, Takei Y, Miyajima T, Mori T, Nakayama T, Takekuma K, Hoshika A
Department of Pediatrics, Tokyo Medical College, Japan.
J Clin Virol. 1998 Jul 24;11(1):67-75. doi: 10.1016/s0928-0197(98)00022-1.
Non-polio enterovirus infections are recognized in children during summer-fall seasons and they sometimes cause large outbreaks. We experienced a nosocomial infection in the neonatal nursery and echovirus type 7 was isolated from samples of four patients.
We diagnosed the horizontal infection of four neonates by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) and the nucleotide sequence.
Total RNA was extracted from clinical isolates, serum samples and cerebrospinal fluid (CSF). We amplified enterovirus genome in the 5'-noncoding region by nested PCR and determined the nucleotide sequences.
Enterovirus genome was detected in all isolates, in the acute-phase sera in all four patients and in the CSF in one patient by the first PCR. By using nested PCR, the genome was detected from convalescent-phase sera in two patients. All enterovirus genome obtained from the nursery outbreak showed the same sequences with 100% homology.
We demonstrated the clinical advantages of RT-nested PCR from serum samples and the analysis of nucleotide sequencing gave the supportive evidence of identification of transmission pathway.
非脊髓灰质炎肠道病毒感染在夏秋季节的儿童中较为常见，有时会引发大规模疫情。我们在新生儿重症监护室经历了一次医院感染，并从四名患者的样本中分离出了7型艾柯病毒。
我们通过逆转录巢式聚合酶链反应（RT-nested PCR）和核苷酸序列诊断了四名新生儿的水平感染。
从临床分离株、血清样本和脑脊液（CSF）中提取总RNA。我们通过巢式PCR扩增5'-非编码区的肠道病毒基因组，并确定核苷酸序列。
通过首次PCR，在所有分离株、所有四名患者的急性期血清以及一名患者的脑脊液中均检测到肠道病毒基因组。通过巢式PCR，在两名患者的恢复期血清中检测到了基因组。从新生儿重症监护室疫情中获得的所有肠道病毒基因组显示出相同的序列，同源性为100%。
我们证明了RT-nested PCR在血清样本中的临床优势，核苷酸序列分析为传播途径的鉴定提供了支持性证据。
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