Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections
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 I...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2011-01-01
|
| Series: | Advances in Virology |
| Online Access: | http://dx.doi.org/10.1155/2011/514681 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832558462724210688 |
|---|---|
| author | 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 |
| author_facet | 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 |
| author_sort | Sandra Menting |
| collection | DOAJ |
| description | 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 (R2=0.9967) and a LOD of at least 1.95×104 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. |
| format | Article |
| id | doaj-art-852aa72fc2db488499464c9662e32170 |
| institution | Kabale University |
| issn | 1687-8639 1687-8647 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advances in Virology |
| spelling | doaj-art-852aa72fc2db488499464c9662e321702025-02-03T01:32:22ZengWileyAdvances in Virology1687-86391687-86472011-01-01201110.1155/2011/514681514681Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus InfectionsSandra Menting0Khoa T. D. Thai1Tran T. T. Nga2Hoang L. Phuong3Paul Klatser4Katja C. Wolthers5Tran Q. Binh6Peter J. de Vries7Marcel Beld8Royal Tropical Institute, KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The NetherlandsDivision of Infectious Diseases, Academic Medical Center, Tropical Medicine & AIDS, Meibergdreef 9, 1105 AZ Amsterdam, The NetherlandsDivision of Microbiology, Cho Ray Hospital, 201B Nguyen Chi Thanh Street, District 5, Ho Chi Minh City, VietnamDivision of Infectious Diseases, Academic Medical Center, Tropical Medicine & AIDS, Meibergdreef 9, 1105 AZ Amsterdam, The NetherlandsRoyal Tropical Institute, KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The NetherlandsDivision of Medical Microbiology, Academic Medical Center, Section of Clinical Virology, Meibergdreef 9, 1105 AZ Amsterdam, The NetherlandsDivision of Microbiology, Cho Ray Hospital, 201B Nguyen Chi Thanh Street, District 5, Ho Chi Minh City, VietnamDivision of Infectious Diseases, Academic Medical Center, Tropical Medicine & AIDS, Meibergdreef 9, 1105 AZ Amsterdam, The NetherlandsRoyal Tropical Institute, KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The NetherlandsDengue 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 (R2=0.9967) and a LOD of at least 1.95×104 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.http://dx.doi.org/10.1155/2011/514681 |
| spellingShingle | 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 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 |
| title | Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections |
| title_full | Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections |
| title_fullStr | Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections |
| title_full_unstemmed | Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections |
| title_short | Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections |
| title_sort | internally controlled generic real time pcr for quantification and multiplex real time pcr with serotype specific probes for serotyping of dengue virus infections |
| url | http://dx.doi.org/10.1155/2011/514681 |
| work_keys_str_mv | AT sandramenting internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT khoatdthai internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT tranttnga internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT hoanglphuong internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT paulklatser internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT katjacwolthers internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT tranqbinh internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT peterjdevries internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections AT marcelbeld internallycontrolledgenericrealtimepcrforquantificationandmultiplexrealtimepcrwithserotypespecificprobesforserotypingofdenguevirusinfections |