Validation of an internally controlled one-step real-time multiplex RT-PCR assay for the detection and quantitation of dengue virus RNA in plasma

被引：89

作者：

Kien Duong Thi Hue ^{[1
]}

Trung Vu Tuan ^{[1
]}

Hanh Tien Nguyen Thi ^{[1
]}

Chau Tran Nguyen Bich ^{[1
]}

Huy Huynh Le Anh ^{[1
]}

Wills, Bridget A. ^{[1
,3
]}

Simmons, Cameron P. ^{[1
,2
]}

机构：

[1] Univ Oxford, Clin Res Unit, Hosp Trop Dis, Ho Chi Minh City, Vietnam

[2] Univ Oxford, Nuffield Dept Clin Med, Ctr Trop Med, Ctr Clin Vaccinol & Trop Med, Oxford, England

[3] Univ Oxford, Churchill Hosp, CCVTM, Ctr Trop Med, Oxford, England

来源：

JOURNAL OF VIROLOGICAL METHODS | 2011年 / 177卷 / 02期

基金：

英国惠康基金;

关键词：

Dengue virus; One step real time multiplex RT-PCR; Validation; RAPID TESTS; ANTIBODY; VIREMIA; SPECIFICITY; SENSITIVITY; INFECTION; DIAGNOSIS; PHASE;

D O I：

10.1016/j.jviromet.2011.08.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Dengue is mosquito-borne virus infection that annually causes similar to 50 million clinically apparent cases worldwide. An internally controlled one-step real-time multiplex RT-PCR assay was developed for detection and quantitation of DENV RNA in plasma sample by using specific primers and fluorogenic TaqMan probes. All primers and probes targeted sequences near the 3' end of the NS5 gene. The method comprised two multiplex assays and was validated for sensitivity, specificity, linearity, reproducibility and precision. An internal control template was spiked into each clinical specimen to provide quality assurance for each experimental step. The assay allowed for detection of between 0.5 and 3 infectious particles per mL, is rapid and has been operationally characterized in 287 Vietnamese dengue patients from two therapeutic intervention trials at the Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：168 / 173

页数：6

共 26 条

[1] Epidemiological Factors Associated with Dengue Shock Syndrome and Mortality in Hospitalized Dengue Patients in Ho Chi Minh City, Vietnam [J].

Anders, Katherine L. ;

Nguyen Minh Nguyet ;

Nguyen Van Vinh Chau ;

Nguyen Thanh Hung ;

Tran Thi Thuy ;

Le Bich Lien ;

Jeremy Farrar ;

Bridget Wills ;

Tran Tinh Hien ;

Simmons, Cameron P. .

AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 2011, 84 (01) :127-134

[2]

[Anonymous], J INFECT DIS

[3]

[Anonymous], [No title captured]

[4] Development of real-time reverse transcriptase PCR assays to detect and serotype dengue viruses [J].

Chien, LJ ;

Liao, TL ;

Shu, PY ;

Huang, JH ;

Gubler, DJ ;

Chang, GJJ .

JOURNAL OF CLINICAL MICROBIOLOGY, 2006, 44 (04) :1295-1304

[5] 2nd International External Quality Control Assessment for the Molecular Diagnosis of Dengue Infections [J].

Domingo, Cristina ;

Niedrig, Matthias ;

Teichmann, Anette ;

Kaiser, Marco ;

Rumer, Leonid ;

Jarman, Richard G. ;

Donoso-Mantke, Oliver .

PLOS NEGLECTED TROPICAL DISEASES, 2010, 4 (10)

[6] Evaluation of Two New Commercial Tests for the Diagnosis of Acute Dengue Virus Infection Using NS1 Antigen Detection in Human Serum [J].

Dussart, Philippe ;

Petit, Laure ;

Labeau, Bhety ;

Bremand, Laetitia ;

Leduc, Alexandre ;

Moua, David ;

Matheus, Severine ;

Baril, Laurence .

PLOS NEGLECTED TROPICAL DISEASES, 2008, 2 (08)

[7] Multi-Country Evaluation of the Sensitivity and Specificity of Two Commercially-Available NS1 ELISA Assays for Dengue Diagnosis [J].

Guzman, Maria G. ;

Jaenisch, Thomas ;

Gaczkowski, Roger ;

Vo Thi Ty Hang ;

Sekaran, Shamala Devi ;

Kroeger, Axel ;

Vazquez, Susana ;

Ruiz, Didye ;

Martinez, Eric ;

Mercado, Juan C. ;

Balmaseda, Angel ;

Harris, Eva ;

Dimano, Efren ;

Leano, Prisca Susan A. ;

Yoksan, Sutee ;

Villegas, Elci ;

Benduzu, Herminia ;

Villalobos, Iris ;

Farrar, Jeremy ;

Simmons, Cameron P. .

PLOS NEGLECTED TROPICAL DISEASES, 2010, 4 (08)

[8] Diagnostic Accuracy of NS1 ELISA and Lateral Flow Rapid Tests for Dengue Sensitivity, Specificity and Relationship to Viraemia and Antibody Responses [J].

Hang, Vu Ty ;

Nguyet, Nguyen Minh ;

Trung, Dinh The ;

Tricou, Vianney ;

Yoksan, Sutee ;

Dung, Nguyen Minh ;

Van Ngoc, Tran ;

Hien, Tran Tinh ;

Farrar, Jeremy ;

Wills, Bridget ;

Simmons, Cameron P. .

PLOS NEGLECTED TROPICAL DISEASES, 2009, 3 (01)

[9] Kinetics of non-structural protein 1, IgM and IgG antibodies in dengue type 1 primary infection [J].

Hu, Dongmei ;

Di, Biao ;

Ding, Xixia ;

Wang, Yadi ;

Chen, Yue ;

Pan, Yuxian ;

Wen, Kun ;

Wang, Ming ;

Che, Xiaoyan .

VIROLOGY JOURNAL, 2011, 8

[10]

*ICH, 1995, INT COOP HARM TECHN

← 1 2 3 →