Microchip, reverse transcription‐polymerase chain reaction and culture methods to detect enterovirus infection in pediatric patients

Tsao, Lon-Yen; Lin, Chia-Cheng; Yu, Ya-Yan; Wang, Bao-Tyan

doi:10.1111/j.1442-200x.2006.02157.x

Cited by 19 publications

(7 citation statements)

References 11 publications

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“…A recent study directly compared EV-PCR of CSF samples with a microchip method (DR. Chip Biotechnology). The microchip method detected enterovirus in 52% of CSF samples from EV71-infected patients, compared with a detection rate of 29% for CSF samples tested by EV-PCR [30]. All CSF samples that were positive for enterovirus at TCH were also positive by EV-PCR at the CDC laboratory.…”

Section: Discussionmentioning

confidence: 92%

Outbreak of Neurologic Enterovirus Type 71 Disease: A Diagnostic Challenge

Pérez-Vélez

Anderson

Robinson

et al. 2007

Clinical Infectious Diseases

184

114

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Background. Similar to poliovirus, enterovirus type 71 (EV71) causes severe disease, including aseptic meningitis, encephalitis, acute flaccid paralysis, and acute cardiopulmonary dysfunction. Large epidemics of EV71 infection have been reported worldwide.Methods. After recognition of a cluster of cases of EV71 disease, we reviewed records of patients with EV71 disease who required hospitalization at The Children's Hospital in Denver, Colorado, from 2003 through 2005. The presence of enterovirus was detected by reverse-transcriptase polymerase chain reaction (PCR) and/or viral culture of specimens from multiple sources, and the virus was typed as EV71 using genetic sequencing.Results. Eight cases of EV71 disease were identified in both 2003 and 2005. Fifty-six percent of patients with EV71 disease were р6 months of age (range, 4 weeks to 9 years). All 16 patients had EV71 central nervous system infection. Enterovirus PCR (EV-PCR) of cerebrospinal fluid specimens yielded positive results for only 5 (31.2%) of the 16 patients; all of these patients were !4 months of age and had less severe disease. However, EV-PCR of upper respiratory tract specimens yielded positive results for 8 (100%) of 8 patients, and EV-PCR of lower gastrointestinal tract specimens yielded positive results for 7 (87.5%) of 8 patients.Conclusions. An outbreak of neurologic EV71 disease occurred in Denver, Colorado, during 2003 and 2005. Likely, EV71 disease remains unrecognized in other parts of the United States, because EV-PCR of cerebrospinal fluid frequently yields negative results. EV-PCR of specimens from the respiratory and gastrointestinal tracts had higher diagnostic yields than did EV-PCR of cerebrospinal fluid. EV71 infection should be considered in young children presenting with aseptic meningitis, encephalitis, acute flaccid paralysis, or acute cardiopulmonary collapse. EV71 infection may be an underrecognized emerging disease in the United States.

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Section: Discussionmentioning

confidence: 92%

Outbreak of Neurologic Enterovirus Type 71 Disease: A Diagnostic Challenge

Pérez-Vélez

Anderson

Robinson

et al. 2007

Clinical Infectious Diseases

184

114

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“…Most studies did not collect paired sera to measure serotype-specific serum neutralizing antibody in outpatient studies [7], [8], [29]. In our study, paired sera have been collected to verify the validity of the molecular tests.…”

Section: Discussionmentioning

confidence: 99%

“…The traditional standard methods for detection and serotyping of enterovirus infections are virus isolation and immunofluorescence assay (IFA), which are time-consuming and labor-intensive [4], [5]. Several clinical studies have documented that molecular diagnosis based on polymerase chain reaction (PCR) is time-saving and more sensitive than virus isolation for detection of enterovirus infections in hospitalized patients [6]–[7] [8]–[10] but few studies have been conducted in outpatients. Moreover, no study has compared molecular tests and virus isolation/IFA for serotyping of human enteroviruses using clinical specimens.…”

Section: Introductionmentioning

confidence: 99%

Comparing Molecular Methods for Early Detection and Serotyping of Enteroviruses in Throat Swabs of Pediatric Patients

et al. 2012

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BackgroundEnteroviruses include over 100 serotypes and usually cause self-limited infections with non-specific symptoms in children, with the exceptions of polioviruses and enterovirus 71 which frequently cause neurologic complications. Therefore, early detection and serotyping of enteroviruses are critical in clinical management and disease surveillance. Traditional methods for detection and serotyping of enteroviruses are virus isolation and immunofluorescence assay, which are time-consuming. In this study, we compare virus isolation and two molecular tests for detection and serotyping of enteroviruses in clinical samples.MethodsOne hundred and ten throat swabs were collected from pediatric outpatients with enterovirus-like illnesses (hand-foot-mouth disease, herpangina, and non-specific febrile illness). Virus isolation was conducted using multiple cell lines and isolated viruses were serotyped using immunofluorescent assay. In the molecular tests, a semi-nested RT-PCR and a novel CODEHOP platform were used to detect the 5′UTR and VP1 genes of enteroviruses, respectively. Amplified nucleotides were sequenced and genotyped.ResultsAmong the 110 cases, 39(35%), 52(47%), and 46(42%) were tested positive with these three tests, respectively. Using the consensus results of these three tests as the gold standard, agreement of the VP1 CODEHOP test was 96%, which is higher than those of the virus isolation (89%) and the 5′-UTR test (88%). The VP1 CODEHOP test also has the best performance on serotyping confirmed with serum neutralization tests.ConclusionsThe VP1 CODEHOP test performed well for detection and serotyping of enteroviruses in clinical specimens and could reduce unnecessary hospitalization cares during enterovirus seasons.

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“…However, the high prevalence of enteroviruses and serological cross‐reactivity limits clinical and epidemiological utility. Several studies in the last decade have described that molecular diagnosis based on the polymerase chain reaction (PCR) is a more rapid, specific and sensitive approach for the detection and typing of enterovirus infections than virus isolation . Thus, more recently, clinical laboratory diagnosis is generally accomplished by direct molecular testing of fresh clinical specimens to detect enteroviruses .…”

Section: Diagnosismentioning

confidence: 99%

Tissue tropism, pathology and pathogenesis of enterovirus infection

Muehlenbachs

Bhatnagar

Zaki

2014

The Journal of Pathology

159

121

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Enteroviruses are very common and cause infections with a diverse array of clinical features. Enteroviruses are most frequently considered by practising pathologists in cases of aseptic meningitis, encephalitis, myocarditis and disseminated infections in neonates and infants. Congenital infections have been reported and transplacental transmission is thought to occur. Although skin biopsies during hand, foot and mouth disease are infrequently obtained, characteristic dermatopathological findings can be seen. Enteroviruses have been implicated in lower respiratory tract infections. This review highlights histopathological features of enterovirus infection and discusses diagnostic modalities for formalin-fixed paraffin-embedded tissues and their associated pitfalls. Immunohistochemistry can detect enterovirus antigen within cells of affected tissues; however, assays can be non-specific and detect other viruses. Molecular methods are increasingly relied upon but, due to the high frequency of asymptomatic enteroviral infections, clinical-pathological correlation is needed to determine significance. Of note, diagnostic assays on central nervous system or cardiac tissues from immunocompetent patients with prolonged disease courses are most often negative. Histopathological, immunohistochemical and molecular studies performed on clinical specimens also provide insight into enteroviral tissue tropism and pathogenesis.

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Microchip, reverse transcription‐polymerase chain reaction and culture methods to detect enterovirus infection in pediatric patients

Abstract: The DR.EV chip method yielded a statistically higher positive rate and faster test results than the conventional viral culture method.

Cited by 19 publications

References 11 publications

Outbreak of Neurologic Enterovirus Type 71 Disease: A Diagnostic Challenge

Outbreak of Neurologic Enterovirus Type 71 Disease: A Diagnostic Challenge

Comparing Molecular Methods for Early Detection and Serotyping of Enteroviruses in Throat Swabs of Pediatric Patients

Tissue tropism, pathology and pathogenesis of enterovirus infection

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