USEFULNESS OF NESTED PCR AND SEQUENCE-ANALYSIS IN A NOSOCOMIAL OUTBREAK OF NEONATAL ENTEROVIRUS INFECTION

Background: Non-polio enterovirus infections are recognized in childre n during summer-fall seasons and they sometimes cause large outbreaks. We experienced a nosocomial infection in the neonatal nursery and ech ovirus type 7 was isolated from samples of four patients. Objectives: We diagnosed the horizontal infection of four neonates by reverse tran scriptase-nested polymerase chain reaction (RT-nested PCR) and the nuc leotide sequence. Study design: Total RNA was extracted from clinical isolates, serum samples and cerebrospinal fluid (CSF). We amplified en terovirus genome in the 5'-noncoding region by nested PCR and determin ed the nucleotide sequences. Results: Enterovirus genome was detected in all isolates, in the acute-phase sera in all four patients and in t he CSF in one patient by the first PCR. By using nested PCR, the genom e was detected from convalescent-phase sera in two patients. All enter ovirus genome obtained from the nursery outbreak showed the same seque nces with 100% homology. Conclusion: We demonstrated the clinical adva ntages of RT-nested PCR from serum samples and the analysis of nucleot ide sequencing gave the supportive evidence of identification of trans mission pathway. (C) 1998 Elsevier Science B.V. All rights reserved.