Viral diagnosis of neurological infection by RT multiplex PCR: A search for entero- and herpesviruses in a prospective study

Casas, Inmaculada; Pozo, Francisco; Trallero, Gloria; Echevarría, J M; Tenorio, Antonio

doi:10.1002/(sici)1096-9071(199902)57:2<145::aid-jmv10>3.0.co;2-n

Cited by 69 publications

(48 citation statements)

References 34 publications

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“…The incidence of the different viruses varies globally according to different populations, methodologies and objectives of the study. Enteroviruses detected by molecular biology RT-PCR or Real Time PCR or cellular culture analysis show an incidence varying from 10 to 46% 9,12,15,17,19 . Herpesviruses, HHV6, VZV, and a mixed infection by HHV6/EV, were detected in a lower proportion [20][21][22] .…”

Section: Discussionmentioning

confidence: 99%

“…After 2 minutes at 94ºC ice cooling, 17.5 µL of the mix was added to the tube, which was stored at 45ºC for 1 hour. Amplification -PCR Enterovirus -PCR for enterovirus was based on the amplification of the 5'-UTR region of the gene, which is a highly conserved region in most enterovirus serotypes according to Casas et al 9 . For the first PCR the primer sequences were: EV1-Reverse 5'GAAACACG-GACACCAAAGTAGTCG3' and EV1 -Forward 5'CG-GTACCTTTGTRCGCCTGTTTTA3' .…”

Section: Methodsmentioning

confidence: 99%

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Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Vidal

Almeida²,

Messias-Reason³

et al. 2011

Arq. Neuro-Psiquiatr.

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Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8%) samples were positive. Enteroviruses was present in 49 (83%) samples and herpes virus family in 10 (17%), of these 6 (10%) herpes simplex virus, 1 (2%) Epstein Barr virus, 2 (3%) human herpes virus type 6 and 1 (2%) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Vidal

Almeida²,

Messias-Reason³

et al. 2011

Arq. Neuro-Psiquiatr.

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“…Since HSV and enterovirus (EV) are the two most commonly identifiable etiologies of sporadic encephalitis in immunocompetent individuals and since PCR testing for both diseases has become the gold standard for diagnosis (see "HSV" and "EV" sections below), there has been interest in the development of assays that can simultaneously detect these pathogens. Several groups have developed multiplex PCR assays that are capable of detecting HSV-1 and -2, EV, and VZV in a single CSF specimen (36,37,180).…”

Section: Multiplex and Consensus Primer Pcrsmentioning

confidence: 99%

Molecular Methods for Diagnosis of Viral Encephalitis

2004

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Hundreds of viruses cause central nervous system (CNS) disease, including meningoencephalitis and postinfectious encephalomyelitis, in humans. The cerebrospinal fluid (CSF) is abnormal in >90% of cases; however, routine CSF studies only rarely lead to identification of a specific etiologic agent. Diagnosis of viral infections of the CNS has been revolutionized by the advent of new molecular diagnostic technologies to amplify viral nucleic acid from CSF, including PCR, nucleic acid sequence-based amplification, and branched-DNA assay. PCR is ideally suited for identifying fastidious organisms that may be difficult or impossible to culture and has been widely applied for detection of both DNA and RNA viruses in CSF. The technique can be performed rapidly and inexpensively and has become an integral component of diagnostic medical practice in the United States and other developed countries. In addition to its use for identification of etiologic agents of CNS disease in the clinical setting, PCR has also been used to quantitate viral load and monitor duration and adequacy of antiviral drug therapy. PCR has also been applied in the research setting to help discriminate active versus postinfectious immune-mediate disease, identify determinants of drug resistance, and investigate the etiology of neurologic disease of uncertain cause. This review discusses general principles of PCR and reverse transcription-PCR, including qualitative, quantitative, and multiplex techniques, with comment on issues of sensitivity, specificity, and positive and negative predictive values. The application of molecular diagnostic methods for diagnosis of specific infectious entities is reviewed in detail, including viruses for which PCR is of proven efficacy and is widely available, viruses for which PCR is less widely available or for which PCR has unproven sensitivity and specificity, and nonviral entities which can mimic viral CNS disease

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“…In addition, all samples were retested, yielding identical results. The way to avoid false-positive and false-negative results, as well as contamination in the different steps of nested PCR, was described in detail in a previous study carried out in our laboratory (2).…”

Section: Clinical Samples and Patientsmentioning

confidence: 99%

Different Cytomegalovirus Glycoprotein B Genotype Distribution in Serum and Cerebrospinal Fluid Specimens Determined by a Novel Multiplex Nested PCR

2003

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A novel and highly sensitive multiplex nested PCR assay has been developed for the simultaneous glycoprotein B (gB) typing of four gB genotypes of human cytomegalovirus (CMV) directly from clinical specimens. Specifically, a pair of primers to conserved regions of all gB genotypes within the gB gene (gB and gpUL55) was used for primary amplification. A mixture of nested primers to specific and conserved regions of each gB genotype was used for a secondary PCR amplification, yielding amplicons of different sizes for each gB genotype that could easily be differentiated by agarose gel electrophoresis. A total of 40 of 44 serum specimens and 26 of 26 cerebrospinal fluid (CSF) samples, which had previously tested positive for CMV in 66 of 70 AIDS patients with different CMV disease conditions, were gB genotyped by this novel assay. Significant differences in glycoprotein B genotype distribution between serum and CSF specimens were found (P ‫؍‬ 0.001). gB genotype 3 (gB3) and gB2 were the most prevalent genotypes in sera (42.5%) and CSF (38.5%), respectively. A different distribution was also observed when only patients with CMV retinitis were studied (P ‫؍‬ 0.005), suggesting a gB2 neuron tropism. Neither CMV disease nor any particular clinical manifestation of CMV disease was associated with gB genotypes (P > 0.05). A high incidence of mixed infection with the gB1 and gB3 genotypes (27.5%) was detected in serum specimens, indicating that reinfection and reactivation are common traits in advanced AIDS patients.

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Viral diagnosis of neurological infection by RT multiplex PCR: A search for entero- and herpesviruses in a prospective study

Cited by 69 publications

References 34 publications

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Molecular Methods for Diagnosis of Viral Encephalitis

Different Cytomegalovirus Glycoprotein B Genotype Distribution in Serum and Cerebrospinal Fluid Specimens Determined by a Novel Multiplex Nested PCR

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