Duplex Reverse Transcription-PCR Followed by Nested PCR Assays for Detection and Identification of Brazilian Alphaviruses and Flaviviruses

Bronzoni, Roberta Vieira de Morais; Baleotti, Flávia Graciela; Nogueira, Rita Maria Ribeiro; Nunes, Márcio Roberto Teixeira; Figueiredo, Luíz Tadeu Moraes

doi:10.1128/jcm.43.2.696-702.2005

Cited by 159 publications

(137 citation statements)

References 35 publications

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“…RNA extraction was performed with QiAmp Viral RNA (QIAGEN, Germany). RNA was examined using Multiplex-Nested-PCR (M-N-PCR) to detect DENV 1-4 8 . The first RT-PCR was performed using Flavivirus generic primers based on non-structural protein (NS5).…”

Section: Case Reportmentioning

confidence: 99%

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Co-Infection of Dengue Virus by Serotypes 1 and 4 in Patient From Medium Sized City From Brazil

Colombo

Vedovello

Reis

et al. 2013

Rev. Inst. Med. trop. S. Paulo

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Section: Case Reportmentioning

confidence: 99%

“…The Nested-PCR used in our study 8 is a suitable tool for detecting Flavivirus co-infections. Within a short amount of time, the identification of the most common Brazilian Flavivirus is possible.…”

mentioning

confidence: 99%

Co-Infection of Dengue Virus by Serotypes 1 and 4 in Patient From Medium Sized City From Brazil

Colombo

Vedovello

Reis

et al. 2013

Rev. Inst. Med. trop. S. Paulo

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“…The total RNA was reverse transcribed using 100U of reverse transcriptase (Superscript III, Invitrogen), a generic flavivirus primer, 10µL of RNA, and 20U of RNAse inhibitor (RNAse OUT, Invitrogen). The mixture was incubated for 50 min at 50°C and 15 min at 70°C (32) (33) (34) . The cDNA (8µL) was subjected to multiplex semi-nested PCR to detect the DENV serotypes, YFV, WNV, and SLEV (34) .…”

Section: Rna Extraction and Multiplex Semi-nested Reverse Transcriptamentioning

confidence: 99%

“…The mixture was incubated for 50 min at 50°C and 15 min at 70°C (32) (33) (34) . The cDNA (8µL) was subjected to multiplex semi-nested PCR to detect the DENV serotypes, YFV, WNV, and SLEV (34) . Complementary DNA from positive controls was included in the reactions (i.e., DENV-4 strain Boa Vista, SLEV BeH 355964, and YFV 17DD), and a negative control without DNA template (Table 1).…”

Section: Rna Extraction and Multiplex Semi-nested Reverse Transcriptamentioning

confidence: 99%

Natural transovarial transmission of dengue virus 4 in Aedes aegypti from Cuiabá, State of Mato Grosso, Brazil

Cruz

Serra

Santos

et al. 2015

Rev. Soc. Bras. Med. Trop.

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“…Initially, clinical samples were tested for the presence of flavivirus and alphavirus, as described by Bronzoni et al 12 .…”

Section: Study Strategymentioning

confidence: 99%

Concurrent dengue and malaria in the Amazon region

Santana

Lavezzo

Terzian

et al. 2010

Rev. Soc. Bras. Med. Trop.

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INTRODUCTION: The Amazon region has extensive forested areas and natural ecosystems, providing favorable conditions for the existence of innumerous arboviruses. Over 200 arboviruses have been isolated in Brazil and about 40 are associated with human disease. Four out of 40 are considered to be of public health importance in Brazil: Dengue viruses (1-4), Oropouche, Mayaro and Yellow Fever. Along with these viruses, about 98% of the malaria cases are restricted to the Legal Amazon region. METHODS: This study aimed to investigate the presence of arboviruses in 111 clinical serum samples from patients living in Novo Repartimento (Pará), Plácido de Castro (Acre), Porto Velho (Rondônia) and Oiapoque (Amapá). The viral RNA was extracted and RT-PCR was performed followed by a Multiplex-Nested-PCR, using Flavivirus, Alphavirus and Orthobunyavirus generic and species-specific primers. RESULTS: Dengue virus serotype 2 was detected in two patients living in Novo Repartimento (Pará) that also presented active Plasmodium vivax infection. CONCLUSIONS: Despite scant data, this situation is likely to occur more frequently than detected in the Amazon region. Finally, it is important to remember that both diseases have similar clinical findings, thus the diagnosis could be made concomitantly for dengue and malaria in patients living or returning from areas where both diseases are endemic or during dengue outbreaks.

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Duplex Reverse Transcription-PCR Followed by Nested PCR Assays for Detection and Identification of Brazilian Alphaviruses and Flaviviruses

Cited by 159 publications

References 35 publications

Co-Infection of Dengue Virus by Serotypes 1 and 4 in Patient From Medium Sized City From Brazil

Co-Infection of Dengue Virus by Serotypes 1 and 4 in Patient From Medium Sized City From Brazil

Natural transovarial transmission of dengue virus 4 in Aedes aegypti from Cuiabá, State of Mato Grosso, Brazil

Concurrent dengue and malaria in the Amazon region

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