Usefulness of nested PCR and sequence analysis in a nosocomial outbreak of neonatal enterovirus infection.
BACKGROUND
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.


OBJECTIVE
We diagnosed the horizontal infection of four neonates by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) and the nucleotide sequence.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Pediatrics, Tokyo Medical College, Japan.
T
T
Takami
H
H
Kawashima
Y
Y
Takei
T
T
Miyajima
T
T
Mori
T
T
Nakayama
K
K
Takekuma
A
A
Hoshika
D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D002555:Cerebrospinal Fluid / Q000821:virology; D003428:Cross Infection; D018076:DNA, Complementary; D004196:Disease Outbreaks; D004457:Echovirus Infections / Q000453:epidemiology / Q000821:virology*; D029822:Enterovirus B, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007231:Infant, Newborn; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D009725:Nurseries, Hospital; D010614:Pharynx / Q000821:virology; D012367:RNA, Viral / Q000097:blood; D012007:Rectum / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D020121:5' Untranslated Regions; D018076:DNA, Complementary; D012367:RNA, Viral