Internally controlled, generic real-time PCR for quantification and multiplex real-time PCR with serotype-specific probes for serotyping of dengue virus infections

Sandra Menting; Khoa T. D. Thai; Tran T. T. Nga; Hoang L. Phuong; Paul Klatser; Katja C. Wolthers; Tran Q. Binh; Peter J. de Vries; Marcel Beld

doi:https://doi.org/10.1155/2011/514681

Internally controlled, generic real-time PCR for quantification and multiplex real-time PCR with serotype-specific probes for serotyping of dengue virus infections

Sandra Menting, Khoa T. D. Thai, Tran T. T. Nga, Hoang L. Phuong, Paul Klatser, Katja C. Wolthers, Tran Q. Binh, Peter J. de Vries, Marcel Beld

Research output: Contribution to journal › Article › Academic › peer-review

16 Citations (Scopus)

Abstract

Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity (R(2) = 0.9967) and a LOD of at least 1.95 × 10(4) copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping

Original language	English
Pages (from-to)	514681
Journal	Advances in virology
Volume	2011
DOIs	https://doi.org/10.1155/2011/514681
Publication status	Published - 2011

Access to Document

https://doi.org/10.1155/2011/514681

Cite this

Menting, S., Thai, K. T. D., Nga, T. T. T., Phuong, H. L., Klatser, P., Wolthers, K. C., Binh, T. Q., de Vries, P. J., & Beld, M. (2011). Internally controlled, generic real-time PCR for quantification and multiplex real-time PCR with serotype-specific probes for serotyping of dengue virus infections. Advances in virology, 2011, 514681. https://doi.org/10.1155/2011/514681

@article{22a0828c3ae5468c889e2421a9136232,

title = "Internally controlled, generic real-time PCR for quantification and multiplex real-time PCR with serotype-specific probes for serotyping of dengue virus infections",

abstract = "Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity (R(2) = 0.9967) and a LOD of at least 1.95 × 10(4) copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping",

author = "Sandra Menting and Thai, {Khoa T. D.} and Nga, {Tran T. T.} and Phuong, {Hoang L.} and Paul Klatser and Wolthers, {Katja C.} and Binh, {Tran Q.} and {de Vries}, {Peter J.} and Marcel Beld",

year = "2011",

doi = "https://doi.org/10.1155/2011/514681",

language = "English",

volume = "2011",

pages = "514681",

journal = "Advances in virology",

issn = "1687-8647",

publisher = "Hindawi Publishing Corporation",

}

TY - JOUR

T1 - Internally controlled, generic real-time PCR for quantification and multiplex real-time PCR with serotype-specific probes for serotyping of dengue virus infections

AU - Menting, Sandra

AU - Thai, Khoa T. D.

AU - Nga, Tran T. T.

AU - Phuong, Hoang L.

AU - Klatser, Paul

AU - Wolthers, Katja C.

AU - Binh, Tran Q.

AU - de Vries, Peter J.

AU - Beld, Marcel

PY - 2011

Y1 - 2011

N2 - Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity (R(2) = 0.9967) and a LOD of at least 1.95 × 10(4) copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping

AB - Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity (R(2) = 0.9967) and a LOD of at least 1.95 × 10(4) copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping

U2 - https://doi.org/10.1155/2011/514681

DO - https://doi.org/10.1155/2011/514681

M3 - Article

C2 - 22312344

SN - 1687-8647

VL - 2011

SP - 514681

JO - Advances in virology

JF - Advances in virology

ER -