Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay

Dierssen, U.; Rehren, Fabienne; Henke-Gendo, Cornelia; Harste, Gabi; Heim, Albert

doi:10.1016/j.jcv.2007.11.016

Cited by 95 publications

(72 citation statements)

References 38 publications

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“…Real-time PCR was performed as previously described by Dierssen et al with modification (25). Viral RNA was assayed in a 20-l reaction mixture containing 5 l of viral RNA by using the one-step SYBR PrimeScript Plus reverse transcription-PCR (RT-PCR) kit (TaKaRa) with primers EQ-1 and EQ-2 (25). Viral RNA of PV1(Sabin) was used to control the quantification of the number of copies.…”

Section: Methodsmentioning

confidence: 99%

Phosphatidylinositol 4-Kinase III Beta Is a Target of Enviroxime-Like Compounds for Antipoliovirus Activity

Arita

Kojima

Nagano

et al. 2011

J Virol

142

195

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Enviroxime is an antienterovirus compound that targets viral protein 3A and/or 3AB and suppresses a step in enterovirus replication by unknown mechanism. To date, four antienterovirus compounds, i.e., GW5074, Flt3 inhibitor II, TTP-8307, and AN-12-H5, are known to have similar mutations in the 3A protein-encoding region causing resistance to enviroxime (a G5318A [3A-Ala70Thr] mutation in poliovirus [PV]) and are considered enviroxime-like compounds. Recently, antienterovirus activity of a phosphatidylinositol 4-kinase III beta (PI4KB) inhibitor, PIK93, was reported, suggesting that PI4KB is an important host factor targetable by antienterovirus compounds (N. Y. Hsu et al., Cell 141:799-811, 2010). In this study, we analyzed the inhibitory effects of previously identified enviroxime-like compounds (GW5074 and AN-12-H5) and a newly identified antienterovirus compound, T-00127-HEV1, on phosphoinositide (PI) kinases. We found that T-00127-HEV1 inhibited PI4KB activity with a higher specificity for than other PI kinases, in contrast to GW5074, which had a broad specificity for PI kinases. In contrast, AN-12-H5 showed no inhibitory effect on PI4KB activity and only moderate inhibitory effects on PI 3-kinase activity. Small interfering RNA (siRNA) screening targeting PI kinases identified PI4KB is a target of GW5074 and T-00127-HEV1, but not of AN-12-H5, for anti-PV activity. Interestingly, T-00127-HEV1 and GW5074 did not inhibit hepatitis C virus (HCV) replication, in contrast to a strong inhibitory effect of AN-12-H5. These results suggested that PI4KB is an enterovirus-specific host factor required for the replication process and targeted by some enviroxime-like compounds (T-00127-HEV1 and GW5074) and that enviroxime-like compounds may have targets other than PI kinases for their antiviral effect.

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Section: Methodsmentioning

confidence: 99%

Phosphatidylinositol 4-Kinase III Beta Is a Target of Enviroxime-Like Compounds for Antipoliovirus Activity

Arita

Kojima

Nagano

et al. 2011

J Virol

142

195

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“…Previous evaluations of the EV primers and probe in a nonmultiplexed one-step RT-PCR format demonstrated a sensitivity of 100 copies of EV RNA per reaction, corresponding to approximately 3,800 copies/ml of CSF (7). Nonmultiplexed detection for HPeV by one-step real-time RT-PCR showed a 100% detection limit of 30 RNA copies per reaction, derived from testing 30 replicates (19).…”

Section: Discussionmentioning

confidence: 99%

“…For EV amplification, primers and probe (5Ј-TCC-GGC-CCC-TGA-ATG-CGG-CTA-AT-3Ј) were as described by Dierssen et al (7). Primers and probe [5Ј-AAA-CAC-TAG-TTG-TA(A/T)-GGC-CC-3Ј] for HPeV corresponded to those described by Benschop et al (3).…”

Section: In Vitro-transcribed Rna For Species a To D Enteroviruses Anmentioning

confidence: 99%

“…HPeVs in particular historically have been problematic to detect by virus culture and cannot be detected by routine enterovirus RT-PCR; thus, HPeV infections long have been underdiagnosed (2). More recently, separate real-time RT-PCR assays for the detection of EV and HPeV have been developed (3,7,17,19,20); the rapid molecular testing provided by these assays for hospitalized children is important, since it has been shown to reduce antibiotic usage, unnecessary investigations, and duration of hospital stay (3,17,26). In the current study, we have multiplexed EV and HPeV into a single, one-step real-time RT-PCR using two assay formats generally used in two diagnostic virology laboratories for the direct testing of clinical specimens.…”

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confidence: 99%

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Rapid Simultaneous Detection of Enterovirus and Parechovirus RNAs in Clinical Samples by One-Step Real-Time Reverse Transcription-PCR Assay

Bennett¹,

Harvala

Witteveldt

et al. 2011

J Clin Microbiol

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“…Enteroviruses (EVs) are RNA viruses belonging to the Picornaviridae family and are classified into 5 human pathogenic species including poliovirus and human enteroviruses (HEVs) A, B, C and D (Dierssen et al, 2008). Echoviruses and coxsackieviruses are the most commonly identified causes of viral meningitis in the paediatric population (Archimbaud et al, 2004) and in adults (Hong et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Real-time PCR quantitative assessment of hepatitis A virus, rotaviruses and enteroviruses in the Tyume River located in the Eastern Cape Province, South Africa

Sibanda¹,

Okoh²

2013

WSA

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We applied real-time RT-PCR (reverse transcription-polymerase chain reaction) to assess the incidence of hepatitis A virus, rotaviruses and enteroviruses in the Tyume River, an important water resource in the impoverished Eastern Cape Province of South Africa. Detection of noroviruses was done using conventional semi-nested RT-PCR. Water samples were collected once monthly from 6 sampling sites over a 12-month period starting in August 2010 and ending in July 2011. Hepatitis A virus was detected in 13% of the samples in concentrations ranging between 1.67×10 3 genome copies/ℓ and 1.64×10 4 genome copies/ℓ while rotaviruses were detected in 4% of the samples with concentrations ranging from 9×10 1 genome copies/ℓ to 5.64×10 3 genome copies/ℓ. Enteroviruses were not detected in any of the samples, while noroviruses were detected in 4% of the samples. All hepatitis A and rotaviruses positive samples were from the upstream sections of Tyume River while noroviruses were detected in samples from downstream sections only. Statistical analysis showed that occurrence of the viruses in Tyume River was sporadic. Risk analysis showed that hepatitis A virus posed greater risk than rotaviruses for both recreational and domestic water uses. Because of the low infectious dose of enteric viruses, the detection of even low concentrations of hepatitis A virus, rotaviruses and noroviruses in surface water poses a significant risk to public health.

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Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay

Cited by 95 publications

References 38 publications

Phosphatidylinositol 4-Kinase III Beta Is a Target of Enviroxime-Like Compounds for Antipoliovirus Activity

Phosphatidylinositol 4-Kinase III Beta Is a Target of Enviroxime-Like Compounds for Antipoliovirus Activity

Rapid Simultaneous Detection of Enterovirus and Parechovirus RNAs in Clinical Samples by One-Step Real-Time Reverse Transcription-PCR Assay

Real-time PCR quantitative assessment of hepatitis A virus, rotaviruses and enteroviruses in the Tyume River located in the Eastern Cape Province, South Africa

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