Pitfalls in the Diagnosis of Enteroviral Infection in Young Children

Faden, Howard; Patel, Pravin H; Campagna, Linda

doi:10.1097/01.inf.0000226842.23106.a2

Cited by 10 publications

(15 citation statements)

References 13 publications

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“…Accordingly, 15.8% of the culturing‐positive samples were identified with a given serotype. Similar results were presented by Faden et al [2006], when only one of 31 culturing‐positive samples was confirmed positive. The methods used by Faden et al [2006] differ partly from ours but the difficulties in the identification may be the same.…”

Section: Discussionsupporting

confidence: 89%

“…Similar results were presented by Faden et al [2006], when only one of 31 culturing‐positive samples was confirmed positive. The methods used by Faden et al [2006] differ partly from ours but the difficulties in the identification may be the same. There are toxic compounds, for example toxin produced by Clostridium difficile , in stool which can cause effect similar to enterovirus CPE [Schmidt et al, 1980; Faden et al, 2006].…”

Section: Discussionsupporting

confidence: 89%

“…The methods used by Faden et al [2006] differ partly from ours but the difficulties in the identification may be the same. There are toxic compounds, for example toxin produced by Clostridium difficile , in stool which can cause effect similar to enterovirus CPE [Schmidt et al, 1980; Faden et al, 2006]. C. difficile is common in stool of healthy children but it is found rarely in adults [Al‐Jumaili et al, 1984].…”

Section: Discussionmentioning

confidence: 75%

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Early human enterovirus infections in healthy Swedish children participating in the PRODIA pilot study

Simonen-Tikka

Hiekka

Klemola

et al. 2012

Journal of Medical Virology

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Human enteroviruses (HEV) are common, especially in childhood and during the enterovirus season, causing mainly asymptomatic infections but also mild and severe illnesses. Numerous studies have shown the association between HEV infections and type 1 diabetes. Here, the prevalence of HEV infections was studied in healthy Swedish children with increased HLA‐associated risk for type 1 diabetes participating in the PRODIA pilot study in which children were randomized to receive probiotics or placebo during the first 6 months of life. Stool specimens collected from 197 children in every 3 months from the age of 3 to 24 months were screened for HEV using traditional viral culturing method and identified with reverse transcriptase polymerase chain reaction (RT‐PCR) and sequencing of the partial VP1 coding part of the viral genome. Altogether 4.8% (52/1,094) of the specimens were HEV‐positive and 22.3% (44/197) of the children excreted HEV during the follow‐up. HEV‐A and HEV‐B were present in 2.1 and 2.7% of the specimens, respectively. HEV‐C and HEV‐D viruses were not detected. In total, 17 different HEV serotypes were detected and the most common findings were CV‐A9 (13.5%), CV‐A16 (11.5%), and CV‐A2 (9.6%). The majority of the infections (92.3%) were during the enterovirus season extending from July to December. Probiotic treatment did not affect significantly the risk of HEV infections during the 2‐year follow‐up although a trend for transient decrease for HEV positivity (HEV‐A and/or HEV‐B) by the age of 12 months was observed in children who received probiotics [OR 0.40; 95% confidence interval 0.15 to 1.08; P‐value 0.071, generalized estimating (GEE) analysis]. According to the results, HEV‐A findings were nearly as common as HEV‐B findings among the healthy children participating in this study. Also it was shown that serotypes belonging to HEV‐A species can be detected by means of viral culturing. J. Med. Virol. 84:923–930, 2012. © 2012 Wiley Periodicals, Inc.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 75%

See 1 more Smart Citation

Early human enterovirus infections in healthy Swedish children participating in the PRODIA pilot study

Simonen-Tikka

Hiekka

Klemola

et al. 2012

Journal of Medical Virology

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“…Overall, 16.6% (53/320) of the stool samples tested positive using real-time PCR, and the serotypes were identified in 10.6% (34/320) of the total number of samples. This result is higher than in recent reports [13], [14], wherein only 4.8% of the specimens were HEV-positive in Swedish samples. In this study, 21 different enterovirus serotypes were detected, which suggests that enteroviral infections occur primarily in the Asia-Pacific region, and that the combination of cell lines, real-time PCR, and RT-snPCR is more efficient and accurate than traditional PCR that only uses primers 222 and 292.…”

Section: Discussioncontrasting

confidence: 74%

Molecular Identification and Analysis of Human Enteroviruses Isolated from Healthy Children in Shenzhen, China from 2010 to 2011

Chen

et al. 2013

PLoS ONE

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ObjectiveTo determine the prevalence and distribution of human enteroviruses (HEVs) among healthy children in Shenzhen, China.MethodClinical specimens were obtained from 320 healthy children under 5 years old in Shenzhen, China from 2010 to 2011. The specimens were evaluated using real-time PCR and cell cultures. The positive specimens were further tested using reverse transcription-seminested PCR (RT-snPCR). Molecular typing and phylogenetic analysis were based on the sequence determined.ResultsAmong the 320 samples, 34 were tested positive for HEVs (10.6%) and 22 different serotypes were identified using RT-snPCR. PV1 and PV2 were also detected. The predominant serotype observed was EV71 (17.6%), followed by CV-B4 (14.7%). HEV-B was detected most frequently, with an overall prevalence of 47.1%. HEV-A and HEV-C were found in 32.3% and 20.6% of the samples, respectively. No HEV-D was identified. Molecular phylogeny indicated that all EV71 strains were of C4 genotype.ConclusionAlthough a variety of HEVs was detected in healthy children, HEV-B was relatively more prevalent than other HEV species. Considering HEV-A is more prevalent than HEV-B among patients with hand-foot-mouth disease, additional long-term surveillance of HEV is warranted in both asymptomatic and symptomatic populations.

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“…However, the cross-reaction of antibodies to quickly degenerating RD cells was mistaken for positive staining. Faden et al (2006) also had a similar problem with the quickly degenerating cell line A549. The WHO recommends establishing conditions (adjustment of cell count and concentration of fetal calf serum) that will allow cells to last for at least 5-7 days without degeneration (WHO, 2004).…”

Section: Choice Of Cell Linesmentioning

confidence: 99%

A convenient rapid culture assay for the detection of enteroviruses in clinical samples: comparison with conventional cell culture and RT-PCR

Terletskaia-Ladwig¹,

Meier²,

Hahn³

et al. 2008

Journal of Medical Microbiology

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A convenient rapid culture assay (RCA) for the detection of enteroviruses was evaluated against RT-PCR using 576 stool and 102 cerebrospinal fluid (CSF) samples. One hundred and ninety stool samples were also tested by conventional cell culture (CCC). The RCA used immunoperoxidase staining of cell culture plates with a blend of monoclonal antibodies (mAbs) against enterovirus VP1 on the second and sixth days after inoculation. This blend was composed of 5D8/1 (Dako) and four 'in-house' mAbs. CCC was performed using fluorescence staining with the Enterovirus Screening Set (Chemicon International) for culture confirmation. Detection of enteroviruses by the RCA was more successful in colonic carcinoma (CaCo-2) and rhabdomyosarcoma (RD) cells than in human embryonic lung fibroblasts, HEp2 and A549 cells. The performance of CCC in RD cells was hindered by rapid cell degeneration and non-specific staining of cells during culture confirmation. The sensitivity of the RCA compared to RT-PCR in stool samples was found to be 71 % (115/161) on the second day and 87 % (140/161) on the sixth day. The sensitivity of the RCA in CSF samples was 38 % (22/58) after 2 days and 59 % (34/58) after 6 days. The specificity of the RCA was 100 %. All CCC-positive samples were positive by the RCA. CCC required 3-14 days for virus recovery. In conclusion, the RCA has the same sensitivity as CCC, significantly shortens the time required for the detection of enteroviruses, and prevents pitfalls associated with using RD cells for CCC. For diagnosis of aseptic meningitis in CSF samples, RT-PCR should be performed.

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Pitfalls in the Diagnosis of Enteroviral Infection in Young Children

Cited by 10 publications

References 13 publications

Early human enterovirus infections in healthy Swedish children participating in the PRODIA pilot study

Early human enterovirus infections in healthy Swedish children participating in the PRODIA pilot study

Molecular Identification and Analysis of Human Enteroviruses Isolated from Healthy Children in Shenzhen, China from 2010 to 2011

A convenient rapid culture assay for the detection of enteroviruses in clinical samples: comparison with conventional cell culture and RT-PCR

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