Resequencing microarray method for molecular diagnosis of human arboviral diseases

N. Berthet, S. Paulous, Lark L Schneider, M. P. Frenkiel, I. Moltini, C. Tran, S. Matheus, C. Ottone, M. N. Ungeheuer, C. Renaudat, V. Caro, P. Dussart, A. Gessain, P. Desprès

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences. It can be used for detecting emerging viruses by revealing the nucleotide polymorphisms within the target of interest. Objectives/study design: As a new tool for molecular diagnosis of arbovirus infection, high density PathogenID v2.0 RMA (PID2-RMA) was assessed for the detection and genetic analysis of dengue, West Nile, and Chikungunya viruses in spiked blood samples or sera from individuals infected with dengue virus. Viral RNAs extracted from biological samples were retrotranscribed into cDNA and amplified using the Phi 29 polymerase-based method. This amplified cDNA was used for hybridization on PID2-RMA. Results: A good specificity of RMA-based detection was demonstrated using a panel of arboviruses including Dengue, West Nile and Chikungunya viruses. This technology was also efficient for the detection and genetic analysis of the different serotypes of dengue virus in sera of infected patients. Furthermore, the mixing of dengue, West Nile and Chikungunya prototype viruses within a single sample of human blood did not interfere with the sensitivity of PID2-RMA. Conclusions: Our data show that high density PID2-RMA was suitable for the identification of medically important arboviruses. It appears to be particularly adapted to the genetic analysis of dengue, West Nile, and Chikungunya viruses in urgent clinical situations where the rapid identification and characterization of the pathogen is essential.

Original languageEnglish (US)
Pages (from-to)238-243
Number of pages6
JournalJournal of Clinical Virology
Volume56
Issue number3
DOIs
StatePublished - Mar 2013
Externally publishedYes

Fingerprint

Chikungunya virus
Dengue
West Nile virus
Oligonucleotide Array Sequence Analysis
Arboviruses
Dengue Virus
Arbovirus Infections
Complementary DNA
Technology
Viral RNA
Serum
Nucleotides
Viruses

Keywords

  • Arboviruses
  • Diagnosis
  • Human blood samples
  • Resequencing microarray

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Resequencing microarray method for molecular diagnosis of human arboviral diseases. / Berthet, N.; Paulous, S.; Schneider, Lark L; Frenkiel, M. P.; Moltini, I.; Tran, C.; Matheus, S.; Ottone, C.; Ungeheuer, M. N.; Renaudat, C.; Caro, V.; Dussart, P.; Gessain, A.; Desprès, P.

In: Journal of Clinical Virology, Vol. 56, No. 3, 03.2013, p. 238-243.

Research output: Contribution to journalArticle

Berthet, N, Paulous, S, Schneider, LL, Frenkiel, MP, Moltini, I, Tran, C, Matheus, S, Ottone, C, Ungeheuer, MN, Renaudat, C, Caro, V, Dussart, P, Gessain, A & Desprès, P 2013, 'Resequencing microarray method for molecular diagnosis of human arboviral diseases', Journal of Clinical Virology, vol. 56, no. 3, pp. 238-243. https://doi.org/10.1016/j.jcv.2012.10.022
Berthet, N. ; Paulous, S. ; Schneider, Lark L ; Frenkiel, M. P. ; Moltini, I. ; Tran, C. ; Matheus, S. ; Ottone, C. ; Ungeheuer, M. N. ; Renaudat, C. ; Caro, V. ; Dussart, P. ; Gessain, A. ; Desprès, P. / Resequencing microarray method for molecular diagnosis of human arboviral diseases. In: Journal of Clinical Virology. 2013 ; Vol. 56, No. 3. pp. 238-243.
@article{5fcb5c32d7a14362b80e9cce4883162d,
title = "Resequencing microarray method for molecular diagnosis of human arboviral diseases",
abstract = "Background: Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences. It can be used for detecting emerging viruses by revealing the nucleotide polymorphisms within the target of interest. Objectives/study design: As a new tool for molecular diagnosis of arbovirus infection, high density PathogenID v2.0 RMA (PID2-RMA) was assessed for the detection and genetic analysis of dengue, West Nile, and Chikungunya viruses in spiked blood samples or sera from individuals infected with dengue virus. Viral RNAs extracted from biological samples were retrotranscribed into cDNA and amplified using the Phi 29 polymerase-based method. This amplified cDNA was used for hybridization on PID2-RMA. Results: A good specificity of RMA-based detection was demonstrated using a panel of arboviruses including Dengue, West Nile and Chikungunya viruses. This technology was also efficient for the detection and genetic analysis of the different serotypes of dengue virus in sera of infected patients. Furthermore, the mixing of dengue, West Nile and Chikungunya prototype viruses within a single sample of human blood did not interfere with the sensitivity of PID2-RMA. Conclusions: Our data show that high density PID2-RMA was suitable for the identification of medically important arboviruses. It appears to be particularly adapted to the genetic analysis of dengue, West Nile, and Chikungunya viruses in urgent clinical situations where the rapid identification and characterization of the pathogen is essential.",
keywords = "Arboviruses, Diagnosis, Human blood samples, Resequencing microarray",
author = "N. Berthet and S. Paulous and Schneider, {Lark L} and Frenkiel, {M. P.} and I. Moltini and C. Tran and S. Matheus and C. Ottone and Ungeheuer, {M. N.} and C. Renaudat and V. Caro and P. Dussart and A. Gessain and P. Despr{\`e}s",
year = "2013",
month = "3",
doi = "10.1016/j.jcv.2012.10.022",
language = "English (US)",
volume = "56",
pages = "238--243",
journal = "Journal of Clinical Virology",
issn = "1386-6532",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Resequencing microarray method for molecular diagnosis of human arboviral diseases

AU - Berthet, N.

AU - Paulous, S.

AU - Schneider, Lark L

AU - Frenkiel, M. P.

AU - Moltini, I.

AU - Tran, C.

AU - Matheus, S.

AU - Ottone, C.

AU - Ungeheuer, M. N.

AU - Renaudat, C.

AU - Caro, V.

AU - Dussart, P.

AU - Gessain, A.

AU - Desprès, P.

PY - 2013/3

Y1 - 2013/3

N2 - Background: Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences. It can be used for detecting emerging viruses by revealing the nucleotide polymorphisms within the target of interest. Objectives/study design: As a new tool for molecular diagnosis of arbovirus infection, high density PathogenID v2.0 RMA (PID2-RMA) was assessed for the detection and genetic analysis of dengue, West Nile, and Chikungunya viruses in spiked blood samples or sera from individuals infected with dengue virus. Viral RNAs extracted from biological samples were retrotranscribed into cDNA and amplified using the Phi 29 polymerase-based method. This amplified cDNA was used for hybridization on PID2-RMA. Results: A good specificity of RMA-based detection was demonstrated using a panel of arboviruses including Dengue, West Nile and Chikungunya viruses. This technology was also efficient for the detection and genetic analysis of the different serotypes of dengue virus in sera of infected patients. Furthermore, the mixing of dengue, West Nile and Chikungunya prototype viruses within a single sample of human blood did not interfere with the sensitivity of PID2-RMA. Conclusions: Our data show that high density PID2-RMA was suitable for the identification of medically important arboviruses. It appears to be particularly adapted to the genetic analysis of dengue, West Nile, and Chikungunya viruses in urgent clinical situations where the rapid identification and characterization of the pathogen is essential.

AB - Background: Resequencing DNA microarray (RMA) technology uses probes designed to identify a panel of viral sequences. It can be used for detecting emerging viruses by revealing the nucleotide polymorphisms within the target of interest. Objectives/study design: As a new tool for molecular diagnosis of arbovirus infection, high density PathogenID v2.0 RMA (PID2-RMA) was assessed for the detection and genetic analysis of dengue, West Nile, and Chikungunya viruses in spiked blood samples or sera from individuals infected with dengue virus. Viral RNAs extracted from biological samples were retrotranscribed into cDNA and amplified using the Phi 29 polymerase-based method. This amplified cDNA was used for hybridization on PID2-RMA. Results: A good specificity of RMA-based detection was demonstrated using a panel of arboviruses including Dengue, West Nile and Chikungunya viruses. This technology was also efficient for the detection and genetic analysis of the different serotypes of dengue virus in sera of infected patients. Furthermore, the mixing of dengue, West Nile and Chikungunya prototype viruses within a single sample of human blood did not interfere with the sensitivity of PID2-RMA. Conclusions: Our data show that high density PID2-RMA was suitable for the identification of medically important arboviruses. It appears to be particularly adapted to the genetic analysis of dengue, West Nile, and Chikungunya viruses in urgent clinical situations where the rapid identification and characterization of the pathogen is essential.

KW - Arboviruses

KW - Diagnosis

KW - Human blood samples

KW - Resequencing microarray

UR - http://www.scopus.com/inward/record.url?scp=84872847081&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872847081&partnerID=8YFLogxK

U2 - 10.1016/j.jcv.2012.10.022

DO - 10.1016/j.jcv.2012.10.022

M3 - Article

C2 - 23219893

AN - SCOPUS:84872847081

VL - 56

SP - 238

EP - 243

JO - Journal of Clinical Virology

JF - Journal of Clinical Virology

SN - 1386-6532

IS - 3

ER -