Sensitivity of rhabdomyosarcoma and guinea pig embryo cell cultures to field isolates of difficult-to-cultivate group A coxsackieviruses

Lipson, Steven M.; Walderman, R; Costello, Patrick; Szabo, Katarina

doi:10.1128/jcm.26.7.1298-1303.1988

Cited by 61 publications

(28 citation statements)

References 13 publications

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“…Collectively, these viruses are associated with diverse clinical manifestations ranging from mild febrile illness to CNS (aseptic meningitis, encephalitis), myocarditis, neonatal systemic enteroviral disease, and paralytic poliomyelitis (12,320). Recovery of enteroviruses in cell cultures is limited by low sensitivity as well as the poor growth characteristics of many serotypes (279). Rotbart described the utility of PCR methods over cell culture methods for rapidly detecting CNS enterovirus infection (419).…”

Section: Qualitative Viral Assaysmentioning

confidence: 99%

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

et al. 2006

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SUMMARY Real-time PCR has revolutionized the way clinical microbiology laboratories diagnose many human microbial infections. This testing method combines PCR chemistry with fluorescent probe detection of amplified product in the same reaction vessel. In general, both PCR and amplified product detection are completed in an hour or less, which is considerably faster than conventional PCR detection methods. Real-time PCR assays provide sensitivity and specificity equivalent to that of conventional PCR combined with Southern blot analysis, and since amplification and detection steps are performed in the same closed vessel, the risk of releasing amplified nucleic acids into the environment is negligible. The combination of excellent sensitivity and specificity, low contamination risk, and speed has made real-time PCR technology an appealing alternative to culture- or immunoassay-based testing methods for diagnosing many infectious diseases. This review focuses on the application of real-time PCR in the clinical microbiology laboratory.

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Section: Qualitative Viral Assaysmentioning

confidence: 99%

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

et al. 2006

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“…The high prevalence of coxsackieviruses A of HEV-C in the present study is interesting as this may suggest a geographical regional difference with respect to enterovirus serotypes or genotypes distribution. This group of human enterovirus has rarely been detected in both clinical and environmental samples as they grow poorly in a conventional cell culture system (Lipson et al 1988). More recently, an outbreak of meningitis associated with coxsackievirus A1 among a group of travellers after swimming in contaminated seawater in Mexico has been reported (Begier et al 2008).…”

Section: C2-031108mentioning

confidence: 99%

Prevalence and genetic diversity of waterborne pathogenic viruses in surface waters of tropical urban catchments

Gin

2011

Journal of Applied Microbiology

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Aims: To study the virological quality of surface water from highly urbanized tropical water catchment areas and to determine predominant enteric viral genotypes in surface water. Methods and Results: A wide range of human pathogenic viruses in urban surface waters was screened by nested PCR assays after concentration by ultrafiltration. Among the 84 water samples collected, at least one virus was detected in 70 (83·3%) of these samples. Noroviruses were determined to be the most prevalent enteric viruses detected in urban surface water samples, followed by astroviruses, enteroviruses, adenoviruses and hepatitis A viruses. The molecular characterization of environmental viral isolates suggested co‐circulation of multiple genotypes of both noroviruses GI and GII, astroviruses and enteroviruses in urban surface waters. Conclusions: Human enteric viruses with great genetic diversity were detected in surface waters, indicating the presence of human origin of faecal contamination in highly urbanized water catchment areas. Significance and Impact of the Study: The present study identifies and characterizes potential viral hazards of source waters for drinking water supply and recreational activities. This will enable scientific decisions to be made regarding the selection and prioritization of human pathogenic viruses to be included in the future risk assessment and treatment evaluation for water and wastewater.

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“…As no vaccine or antiviral drug is currently available, early and rapid detection is critical for HFMD prevention and con-trol (30). At present, the causative agents of HFMD can be effectively diagnosed by the detection of infectious virus, viral antigens, viral genomic RNA, or antiviral antibodies (11,17,25,27). Due to their speed, sensitivity, and specificity, reverse transcription-PCR (RT-PCR)-based molecular diagnostic assays are increasingly used to detect EV71 and CA16 RNA for HFMD diagnosis (8,14,15).…”

mentioning

confidence: 99%

External Quality Assessment for Enterovirus 71 and Coxsackievirus A16 Detection by Reverse Transcription-PCR Using Armored RNA as a Virus Surrogate

Song

Sun

et al. 2011

J Clin Microbiol

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Three armored RNAs (virus-like particles [VLPs]) containing target sequences from enterovirus 71 (EV71) and coxsackievirus A16 (CA16) and a pan-enterovirus (pan-EV) sequence were constructed and used in an external quality assessment (EQA) to determine the performance of laboratories in the detection of EV71 and CA16. The EQA panel, which consisted of 20 samples, including 14 positive samples with different concentrations of EV and either EV71 or CA16 armored RNAs, 2 samples with all 3 armored RNAs, and 4 negative-control samples (NaN 3 -preserved minimal essential medium [MEM] without VLPs), was distributed to 54 laboratories that perform molecular diagnosis of hand, foot, and mouth disease (HFMD) virus infections. A total of 41 data sets from 41 participants were returned; 5 (12.2%) were generated using conventional in-house reverse transcription-PCR (RT-PCR) assays, and 36 (87.8%) were generated using commercial real-time RT-PCR assays. Performance assessments of laboratories differed; 12 (29.3%) showed a need for improvement. Surprisingly, 4 laboratories were unable to detect EV71 RNA in any samples, even those containing the highest concentration of 10 7 IU/ml. Furthermore, the detection sensitivity for EV71 among all laboratories (82.1%) was substantially lower than that for EV (97.4%) or CA16 (95.1%). Overall, the results of the present study indicate that EQA should be performed periodically to help laboratories monitor their ability to detect HFMD viruses and to improve the comparability of results from different laboratories.

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Sensitivity of rhabdomyosarcoma and guinea pig embryo cell cultures to field isolates of difficult-to-cultivate group A coxsackieviruses

Cited by 61 publications

References 13 publications

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

Prevalence and genetic diversity of waterborne pathogenic viruses in surface waters of tropical urban catchments

External Quality Assessment for Enterovirus 71 and Coxsackievirus A16 Detection by Reverse Transcription-PCR Using Armored RNA as a Virus Surrogate

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