However, the effectiveness of this newly developed mumps virus PCR was evaluated with CSF from only six patients with CNS disease. The sensitivity of the LightCycler assay was compared to that of the conventional assay (10) using titrations of HSV-1, HSV-2, and enterovirus isolates that were supplied by the European Union Concerted Action for Quality Control of Nucleic Acid Amplification in Diagnostic Virology (EU CA) program. Noutsias M, Fechner H, de Jonge H, Wang X, Dekkers D, Houtsmuller AB, Pauschinger M, Bergelson J, Warraich R, Yacoub M, Hetzer R, Lamers J, Schultheiss HP, Poller W. In the second phase, 78 CSF specimens from patients hospitalized for CNS infections that had been previously tested by standardized commercially available PCR and RT-PCR assays for neurotropic virus detection were retrospectively analyzed, assessing the application of this commercially available multiplex RT-PCR DNA microarray on a routine basis. Positivt resultat taler for akut enterovirusinfektion eller parechovirusinfektion. Other molecular methods that assist in serotype-specific identification include multiplex real-time hybridization probe RT-PCR for specific detection and differentiation of enterovirus type 71 (EV71) and CVA16 (28), combined multiplex RT-PCR and microarray assay to detect and differentiate EV71 and CVA16 (4), molecular characterization of PV strains by RT-PCR-restriction fragment length polymorphisms (2), and an integrated micro-RT-PCR system for automatic detection of microorganisms, including EV71 (11). To check the false positive results, we also performed GeneXpert Enterovirus Assay (GXEA) with positive samples from TMC-PCR assay.
Serotyping was achieved from 6 specimens with two polio viruses type 1 (vaccine type), two echoviruses 14, one echovirus 5 and one echovirus 30. RNA dilutions (10-fold) from 10−1 to 10−8 of the EV reference strains EV71, CVB2, E30, CVA24, and P1 were prepared to assess the sensitivity of the RT-PCR assay combined with CLP technology. It also plays an important role in the metabolism of the brain. While servicing our clients’ (your) market research requirements, we keep the same approach in focus to help you get the best value for your $$s. The study included 476 CSF specimens collected from nine different laboratories in five European countries. Alternatively, enterovirus PCR has become standard practice due to its much-improved sensitivity and turnaround time. Evidence for recent EV infection can also be gained from stool culture or demonstration of specific immunoglobulin M antibody (11), but this provides only circumstantial evidence of an etiologic role in current central nervous system (CNS) pathology.
Culture is no longer necessary for clinical diagnosis and should only be performed on PCR-positive samples to obtain isolates for typing purposes. These two individual assays (together with a duplex assay for S. Study co-authors, all from Children’s Hospital and the University of Pennsylvania School of Medicine, are Rebecca L. Glucose levels are normal or mildly decreased, while the protein level is normal or slightly increased. ELISA methods that test for homotypical antibodies are impractical unless there is a clinical suspicion of the presence of one particular serotype, whereas heterotypical antibody assays allow detection of most enterovirus serotypes. Quantification of DNA enables further research of disease prognosis and treatment. The Super E-Mix procedure had greater sensitivity than conventional cell culture, but RT-PCR was the most sensitive technique with this type of specimen.
In comparison, real-time PCR allows same-day detection. Traditional cell culture methods require 6 days, on average, for enterovirus detection. Viral culture is the “gold standard” for the diagnosis of enterovirus infection in different clinical specimens such as feces, nose-throat (NT) swabs, bronchoalveolar lavage (BAL), and cerebrospinal fluid (CSF). Phylogenetic analysis showed that these UK Midlands echovirus 30 VP1 sequences clustered most closely with those from Europe and China. © Wiley-Liss, Inc. Overall, 59% of patients with a positive viral PCR had either CSF or serum raised IFN-alpha. Children, neonates, and immunocompromised individuals are particularly susceptible to enterovirus infection of the central nervous system (2, 16, 22).
Methods involving the amplification of nucleic acids have replaced traditional culture-based methods as the gold standard for the detection of EVs in cerebrospinal fluid (CSF) because of their increased sensitivity and speed. CSF samples with negative results for EV RT-PCR have more erythrocytes. The assay was used to assess the incidence of enterovirus and parechovirus RNA in cell culture negative CSF and throat swab samples (n = 200). The sensitivity and specificity of the RT-PCR assay were tested by using all 64 known EV serotypes, several non-EV serotypes, and two Quality Control for Molecular Diagnostics (QCMD) Program EV proficiency panels from 2001 and 2002. In our infant population, multiple singleplex PCR assays perform better than a multiplex assay for the detection of CSF viruses. This test was compared with another EV IgM capture technique, the solid-phase reverse immunosorbent test (SPRIST). Lack of microbiological documentation results in unnecessary antimicrobial therapy and hospitalization.
A specific 113-bp fragment was amplified using primers designed based on 5′ non coding region of the enterovirus genome.