Rapid Virological Diagnosis of Central Nervous System Infections by Use of a Multiplex Reverse Transcription-PCR DNA Microarray

Lévêque, Nicolas; Haecke, Adrien Van; Renois, Fanny; Boutolleau, David; Talmud, Déborah; Andréoletti, Laurent

doi:10.1128/jcm.01214-11

Cited by 53 publications

(32 citation statements)

References 45 publications

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“…For each sample, 80 ng of total nucleic acid extracts was subjected to RT-qPCR using either commercial (Argene EBV R-gene and HSV1 HSV2 VZV [herpes simplex virus 1 and 2 and varicella-zoster virus] R-gene assays; bioMérieux, Verniolle, France) or in-house techniques for the detection of cytomegalovirus (CMV), parvovirus B19 (PVB19), enterovirus (EV), and adenovirus (ADV) (8,(18)(19)(20)(21)(22)(23). In positive samples, the viral load was expressed as the number of genomic DNA or RNA copies per g of total extracted nucleic acids (DNA and RNA) (6).…”

Section: Patientsmentioning

confidence: 99%

Virus Detection and Semiquantitation in Explanted Heart Tissues of Idiopathic Dilated Cardiomyopathy Adult Patients by Use of PCR Coupled with Mass Spectrometry Analysis

Nguyen¹,

Renois²,

Lévêque³

et al. 2013

J Clin Microbiol

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Section: Patientsmentioning

confidence: 99%

Virus Detection and Semiquantitation in Explanted Heart Tissues of Idiopathic Dilated Cardiomyopathy Adult Patients by Use of PCR Coupled with Mass Spectrometry Analysis

Nguyen¹,

Renois²,

Lévêque³

et al. 2013

J Clin Microbiol

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“…The amplified products are then hybridized with the specific probes on the microarray. The images are finally captured using a chip scanner and analytic software and the results are analyzed [15,16].…”

Section: Discussionmentioning

confidence: 99%

Microarray Multiplex Assay for the Simultaneous Detection and Discrimination of Influenza A and Influenza B Viruses

Tian

et al. 2013

Indian J Microbiol

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In this study, we present a microarray approach for the typing of influenza A and B viruses, and the subtyping of H1 and H3 subtypes. We designed four pairs of specific multiplex RT-PCR primers and eight specific oligonucleotide probes and prepared microarrays to identify the specific subtype of influenza virus. Through amplification and fluorescent marking of the multiplex RT-PCR products on the M gene of influenza A and B viruses and the HA gene of subtypes H1 and H3, the PCR products were hybridized with the microarray, and the results were analyzed using a microarray scanner. The results demonstrate that the chip developed by our research institute can detect influenza A and B viruses specifically and identify the subtypes H1 and H3 at a minimum concentration of 1 9 10 2 copies/lL of viral RNA. We tested 35 clinical samples and our results were identical to other fluorescent methods. The microarray approach developed in this study provides a reliable method for the monitoring and testing of seasonal influenza.

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“…A microarray reader piloted by specific software performs the detection by colorimetric reaction and the interpretation of the results. The lower limit of reproducible detection, as determined by Lévêque et al (11), is 250 copies/ml for CMV, 2,000 copies/ml for EBV, and 500 copies/ml for HHV-6.…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, a larger panel of viruses can be tested simultaneously. Microarray techniques have already been evaluated in routine settings for the diagnosis of central nervous system and respiratory tract viral infections (11)(12)(13)(14)(15)(16). To the best of our knowledge, microarrays have not yet been evaluated prospectively for the detection of Herpesviridae in a cohort of patients undergoing HSCT.…”

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confidence: 99%

Detection of Herpesviridae in Whole Blood by Multiplex PCR DNA-Based Microarray Analysis after Hematopoietic Stem Cell Transplantation

et al. 2014

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e Viral infections are important causes of morbidity and mortality in patients after hematopoietic stem cell transplantation. The monitoring by PCR of Herpesviridae loads in blood samples has become a critical part of posttransplant follow-up, representing mounting costs for the laboratory. In this study, we assessed the clinical performance of the multiplex PCR DNA microarray Clart Entherpex kit for detection of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesvirus 6 (HHV-6) as a screening test for virological follow-up. Two hundred fifty-five blood samples from 16 transplanted patients, prospectively tested by routine PCR assays, were analyzed by microarray. Routine PCR detected single or multiple viruses in 42% and 10% of the samples, respectively. Microarray detected single or multiple viruses in 34% and 18% of the samples, respectively. Microarray results correlated well with CMV and EBV detections by routine PCR (kappa tests ‫؍‬ 0.79 and 0.78, respectively), whereas a weak correlation was observed with HHV-6 (0.43). HHV-7 was also detected in 48 samples by microarray. In conclusion, the microarray is a reliable screening assay for a posttransplant virological follow-up to detect CMV and EBV infections in blood. However, positive samples must be subsequently confirmed and viral loads must be quantified by PCR assays. Limitations were identified regarding HHV-6 detection. Although it is promising, is easy to use as a first-line test, and allows a reduction in the cost of analysis without undue delay in the reporting of the final quantitative result to the clinician, some characteristics of this microarray should be improved, particularly regarding quality control and the targeted virus panel, such that it could then be used as a routine test.

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Rapid Virological Diagnosis of Central Nervous System Infections by Use of a Multiplex Reverse Transcription-PCR DNA Microarray

Cited by 53 publications

References 45 publications

Virus Detection and Semiquantitation in Explanted Heart Tissues of Idiopathic Dilated Cardiomyopathy Adult Patients by Use of PCR Coupled with Mass Spectrometry Analysis

Virus Detection and Semiquantitation in Explanted Heart Tissues of Idiopathic Dilated Cardiomyopathy Adult Patients by Use of PCR Coupled with Mass Spectrometry Analysis

Microarray Multiplex Assay for the Simultaneous Detection and Discrimination of Influenza A and Influenza B Viruses

Detection of Herpesviridae in Whole Blood by Multiplex PCR DNA-Based Microarray Analysis after Hematopoietic Stem Cell Transplantation

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