Semi-nested PCR using NS3 primers for the detection and typing of dengue viruses in clinical serum specimens

Seah, C.L.K.; Chow, Vincent T. K.; Chan, Yee Cheun

doi:10.1016/0928-0197(94)00063-z

Cited by 43 publications

(28 citation statements)

References 16 publications

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“…26,36 The reaction mixture consisted of 10 μL 2X PCR master mix (Promega, Madison, WI), 1 μL (10 μM) each forward primer (5 AATTGTCGACAGAAA AGGAAA), and reverse primer (5 GGCTGGGGTTTGGT ATC), 3 μL from the completed RT-PCR and H 2 O to 20 μL. The protocol was 94 C for 2 min, and then 30 cycles of 94 C for 30 sec, and 55 C for 1 min, with an extension at 72 C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Detection of Dengue Virus NS1 Antigen in Infected Aedes aegypti Using a Commercially Available Kit

Voge¹,

Sánchez-Vargas²,

Blair³

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Abstract. Epidemic dengue has emerged throughout the tropical world. In the continued absence of a vaccine against dengue virus (DENV), mosquito vector surveillance and control programs are essential to reduce human infections. An effective test to detect DENV in infected mosquitoes would be a valuable addition to the surveillance effort. We investigated DENV detection in infected Aedes aegypti using a commercially available DENV non-structural protein 1 (NS1) ELISA kit (Platelia Dengue NS1 Ag), and by reverse transcription-polymerase chain reaction (RT-PCR) and virus isolation assays. The DENV-infected mosquitoes were subjected to field-relevant conditions and assayed individually and pooled with uninfected mosquitoes. Overall, DENV NS1 antigen was detected in 98% of infected mosquitoes/ pools versus 79% for RT-PCR and 29% for virus isolation. Our results indicate that NS1 is an excellent analyte for detection of DENV in Ae. aegypti and that the tested NS1 antigen kit provides a sensitive, rapid, and convenient test for DENV surveillance in mosquitoes.

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Section: Methodsmentioning

confidence: 99%

Detection of Dengue Virus NS1 Antigen in Infected Aedes aegypti Using a Commercially Available Kit

Voge¹,

Sánchez-Vargas²,

Blair³

et al. 2013

The American Society of Tropical Medicine and Hygiene

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“…28 DENV RNA was also amplified from acute-phase specimens and serotypes were identified by using reverse transcription-polymerase chain reaction (RT-PCR). 29,30 A 90% plaque reduction neutralization test (PRNT) was used to detect seroconversions using baseline, serosurvey 1, and serosurvey 2 specimens for persons who provided specimens on at least two of these occasions. 31 A participant was considered to have been infected with DENV during the course of the study if he or she had positive results in one or more of these laboratory tests.…”

Section: Methodsmentioning

confidence: 99%

Towards a Casa Segura: A Consumer Product Study of the Effect of Insecticide-Treated Curtains on Aedes aegypti and Dengue Virus Infections in the Home

Loroño-Pino¹,

García-Rejón²,

Machaín-Williams³

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Abstract. The home, or domicile, is the principal environment for transmission of dengue virus (DENV) between humans and mosquito vectors. Community-wide distribution of insecticide-treated curtains (ITCs), mimicking vector control program-driven interventions, has shown promise to reduce DENV infections. We conducted a Casa Segura consumer product intervention study in Mérida, Mexico to determine the potential to reduce intradomicillary DENV transmission through ITC use in individual homes. Dengue virus infections in mosquitoes and in humans were reduced in homes with ITCs in one of two study subareas. Overall, ITCs reduced intradomicillary DENV transmission; ITC homes were significantly less likely to experience multiple DENV infections in humans than NTC homes. Dengue virus-infected Aedes aegypti females were reduced within the ITC homes where curtain use was highest. Some homes yielded up to nine infected Ae. aegypti females. This study provides insights regarding best practices for Casa Segura interventions to protect homes from intradomicillary DENV transmission.

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“…This was followed by RT-PCR-based DENV amplification using primers targeting the NS3 gene. 42 A second round of semi-nested PCR including the upstream consensus primer and DENV 1-4 serotype-specific primers 43 was used to determine DENV serotype. Amplification products were visualized on a 2% LE agarose gel (Promega Corp., Madison, WI) containing ethidium bromide.…”

Section: Introductionmentioning

confidence: 99%

Mosquito Infestation and Dengue Virus Infection in Aedes aegypti Females in Schools in Mérida, México

García-Rejón¹,

Loroño-Pino²,

Farfán-Ale³

et al. 2011

The American Society of Tropical Medicine and Hygiene

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We determined abundance of Aedes aegypti mosquitoes and presence of dengue virus (DENV) in females collected from schools in Mérida, México, during 2008 and 2009. Backpack aspiration from 24 schools produced 468 females of Ae. aegypti and 1,676 females of another human biter, Culex quinquefasciatus. Ae. aegypti females were collected most commonly from classrooms followed by offices and bathrooms. Of these females, 24.7% were freshly fed. Examination of 118 pools of Ae. aegypti females (total of 415 females) for presence of DENV RNA produced 19 positive pools (16.1%). DENV-infected pools were detected from 11 (45.8%) of 24 schools and came from different room types, including classrooms, offices, and bathrooms. The overall rate of DENV infection per 100 Ae. aegypti females was 4.8. We conclude that schools in Mérida present a risk environment for students, teachers, and other personnel to be exposed to mosquitoes and bites of DENV-infected Ae. aegypti females.

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Semi-nested PCR using NS3 primers for the detection and typing of dengue viruses in clinical serum specimens

Cited by 43 publications

References 16 publications

Detection of Dengue Virus NS1 Antigen in Infected Aedes aegypti Using a Commercially Available Kit

Detection of Dengue Virus NS1 Antigen in Infected Aedes aegypti Using a Commercially Available Kit

Towards a Casa Segura: A Consumer Product Study of the Effect of Insecticide-Treated Curtains on Aedes aegypti and Dengue Virus Infections in the Home

Mosquito Infestation and Dengue Virus Infection in Aedes aegypti Females in Schools in Mérida, México

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