Cost-Effective Real-Time Reverse Transcriptase PCR (RT-PCR) To Screen for Dengue Virus followed by Rapid Single-Tube Multiplex RT-PCR for Serotyping of the Virus

Lai, Yee-Ling; Chung, Youne-Kow; Tan, Hwee-Cheng; Yap, H M; Yap, Grace; Ooi, Eng Eong; Ng, Lee Ching

doi:10.1128/jcm.01258-06

Cited by 141 publications

(157 citation statements)

References 18 publications

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“…Of all assays tested, RT-PCR assay described by Lai et al (2007) demonstrated 100% sensitivity (Fig. 1) and specificity in acute sera collected within the first 3 days of fever.…”

Section: Sensitivity Of Dengue Diagnostic Assaysmentioning

confidence: 99%

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Pok

Lai

Sng

et al. 2010

Vector-Borne and Zoonotic Diseases

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Early and accurate diagnosis of dengue is imperative for disease surveillance, which helps in the control of dengue in endemic countries. In this study, we evaluated the performance of three commercially available dengue nonstructural 1 (NS1) antigen assays (Bio-Rad PlateliaÔ Dengue NS1 Antigen ELISA, PanBio Dengue Early ELISA, and Bio-Rad Dengue NS1 Antigen Strip test) and compared them with reverse-transcription polymerase chain reaction (RT-PCR) and other commercially available serological assays for the diagnosis of dengue. The analysis showed RT-PCR to be the most sensitive and specific (100%) diagnostic method during the first 3 days of fever. The overall sensitivity of dengue NS1 antigen assays within the same period was 81.7%, indicating their potential role as a cost-effective and convenient alternative method to RT-PCR for the diagnosis of dengue fever in a primary healthcare setting. However, reduced sensitivity in detecting secondary dengue infections was one of the drawbacks of dengue NS1 antigen assays. Nonetheless, it remains a useful assay for the early detection of dengue and hence could play an important role in routine surveillance efforts to control dengue outbreaks in Singapore.

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“…Of all assays tested, RT-PCR assay described by Lai et al (2007) demonstrated 100% sensitivity (Fig. 1) and specificity in acute sera collected within the first 3 days of fever.…”

Section: Sensitivity Of Dengue Diagnostic Assaysmentioning

confidence: 99%

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Pok

Lai

Sng

et al. 2010

Vector-Borne and Zoonotic Diseases

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“…A DENV 2-specific one-step quantitative RT-PCR assay targeting a 177-bp region of the NS5 gene (Richardson et al 2006) was performed in a LightCycler 2.0 system using a LightCycler RNA Master SYBR Green Kit I (Roche Diagnostics, GMbH). Real-time quantitative RT-PCR assay conditions follow that of Lai et al 2007. Amplification of the target gene from each individual mosquitoes was compared against a standard curve (Efficiency ¼ 1.814, error ¼ 0.0461) generated from a 10-fold serial dilutions of NS5 DENV 2 RNA standard, and LightCycler software version 4.05 was used for data analysis.…”

Section: Evaluation Of Ns1 For Denv Detection With Laboratory-infectementioning

confidence: 99%

“…A dengue group-specific onestep SYBR Green I-based one-step real-time RT-PCR assay was performed as previously described (Lai et al 2007).…”

Section: Evaluation Of Ns1 For Denv Detection With Laboratory-infectementioning

confidence: 99%

Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

Tan

Wong

et al. 2011

Vector-Borne and Zoonotic Diseases

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Dengue fever is currently one of the most important mosquito-borne diseases that affect humans. With neither vaccines nor treatment available, prevention of the disease relies heavily on surveillance and control of mosquito vectors. In the present study, we have evaluated and showed the potential use of the Dengue NS1 Ag Strip Ò for the detection of dengue virus (DENV) in Aedes aegypti. Initial results showed that the sensitivity of the test kit in detecting DENV in wild-caught mosquitoes is comparable to that of real-time reverse transcriptase-polymerase chain reaction. The detection of naturally infected Ae. aegypti with the NS1 rapid test kit in our dengue cluster investigation further illustrates its potential use for surveillance of DENV in wild mosquito populations. The kit can easily be used in a simple field station, and minimal training is required. The results can be obtained in less than an hour. Employment of the kit in the field could help guide mosquito control operations in the prioritization of resources in controlling the transmission of DENV. In this study the potential of the kit for field surveillance of infected dengue vectors, which are epidemiologically important, has been demonstrated.

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“…Real-time RT-PCR methods have been established as a more rapid and sensitive technique for the detection and quantification of DENV in clinical samples (Drosten et al 2002, Lai et al 2007. In this study, we used quantitative real-time RT-PCR to quantify the DENV-4 viral titre (1.08 x 10 3 copies/mL) from a single Ae.…”

Section: Simplexa™ Dengue Real-time Rt-pcr -mentioning

confidence: 99%

Dengue virus type 4 in Niterói, Rio de Janeiro: the role of molecular techniques in laboratory diagnosis and entomological surveillance

Castro¹,

Nogueira²,

Filippis³

et al. 2012

Mem. Inst. Oswaldo Cruz

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In Niterói, state of Rio de Janeiro, dengue virus type 4 (DENV-4) was isolated for the first time in March 2011. We analysed the laboratory findings of the first cases and evaluated the use of molecular techniques for the detection of DENV-4 in Aedes aegypti that were field-caught. Conventional reverse transcriptase-polymerase chain reaction (RT-PCR) and SimplexaTM Dengue real-time RT-PCR confirmed DENV-4 infection in all cases. Additionally, DENV-4 was confirmed in a female Ae. aegypti with 1.08 x 10³ copies/mL of virus, as determined by quantitative real-time RT-PCR. This is the first time the SimplexaTM Dengue real-time assay has been used for the classification of cases of infection and for entomological investigations. The use of these molecular techniques was shown to be important for the surveillance of dengue in humans and vectors

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Cost-Effective Real-Time Reverse Transcriptase PCR (RT-PCR) To Screen for Dengue Virus followed by Rapid Single-Tube Multiplex RT-PCR for Serotyping of the Virus

Cited by 141 publications

References 18 publications

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

Dengue virus type 4 in Niterói, Rio de Janeiro: the role of molecular techniques in laboratory diagnosis and entomological surveillance

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Cost-Effective Real-Time Reverse Transcriptase PCR (RT-PCR) To Screen for Dengue Virus followed by Rapid Single-Tube Multiplex RT-PCR for Serotyping of the Virus

Cited by 141 publications

References 18 publications

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Evaluation of Nonstructural 1 Antigen Assays for the Diagnosis and Surveillance of Dengue in Singapore

Evaluation of the Dengue NS1 Ag Strip®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)

Dengue virus type 4 in Niterói, Rio de Janeiro: the role of molecular techniques in laboratory diagnosis and entomological surveillance

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Evaluation of the Dengue NS1 Ag Strip^®for Detection of Dengue Virus Antigen inAedes aegypti(Diptera: Culicidae)