Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina

Most molecular protocols for Dengue virus detection described so far are time consuming and cumbersome with mosquito samples. In order to count with a sensitive and specific molecular detection system for monitoring possible Dengue outbreaks and circulating viral serotypes in field-caught Aedes aegy...

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Publicado:	2005
Materias:	Aedes aegypti Dengue RFLP RNA RT-PCR primer RNA amplicon Argentina article controlled study Dengue virus gene sequence mosquito nonhuman restriction fragment length polymorphism reverse transcription polymerase chain reaction RNA extraction RNA purification screening serotyping viral genetics virus detection Aedes Animals Base Sequence Dengue Virus Female Humans Insect Vectors Molecular Sequence Data Polymorphism, Restriction Fragment Length Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Nucleic Acid Dengue virus group
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01874640_v47_n3-4_p82_Liotta http://hdl.handle.net/20.500.12110/paper_01874640_v47_n3-4_p82_Liotta
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_01874640_v47_n3-4_p82_Liotta
record_format	dspace
spelling	paper:paper_01874640_v47_n3-4_p82_Liotta2023-06-08T15:19:40Z Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina Aedes aegypti Dengue RFLP RNA RT-PCR primer RNA Aedes aegypti amplicon Argentina article controlled study Dengue virus gene sequence mosquito nonhuman restriction fragment length polymorphism reverse transcription polymerase chain reaction RNA extraction RNA purification screening serotyping viral genetics virus detection Aedes Animals Argentina Base Sequence Dengue Dengue Virus Female Humans Insect Vectors Molecular Sequence Data Polymorphism, Restriction Fragment Length Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Nucleic Acid Aedes aegypti Dengue virus Dengue virus group Most molecular protocols for Dengue virus detection described so far are time consuming and cumbersome with mosquito samples. In order to count with a sensitive and specific molecular detection system for monitoring possible Dengue outbreaks and circulating viral serotypes in field-caught Aedes aegypti populations from Northeastern Argentina, a RT-PCR and RFLP assay was developed. The original RT-PCR assay proposed by Sudiro et al. for human serum was optimized for mosquito samples. Modifications were done at the RNA extraction-purification and at the thermal profile steps. The generic 230 bp amplicon was validated by RFLP assay and cycle sequencing. Results showed that, due to the generic characteristic of the primers used, certain mosquito genome regions could be co-amplified, making confirmation of the Dengue specific amplicon by RFLP assay a required step. Under these conditions, the proposed method can be employed as a Dengue viral generic screening procedure in Aedes aegypti mosquito samples, giving in our hands an estimated 99.52% of confirmed negatives (207/208 tested samples). 2005 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01874640_v47_n3-4_p82_Liotta http://hdl.handle.net/20.500.12110/paper_01874640_v47_n3-4_p82_Liotta
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Aedes aegypti Dengue RFLP RNA RT-PCR primer RNA Aedes aegypti amplicon Argentina article controlled study Dengue virus gene sequence mosquito nonhuman restriction fragment length polymorphism reverse transcription polymerase chain reaction RNA extraction RNA purification screening serotyping viral genetics virus detection Aedes Animals Argentina Base Sequence Dengue Dengue Virus Female Humans Insect Vectors Molecular Sequence Data Polymorphism, Restriction Fragment Length Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Nucleic Acid Aedes aegypti Dengue virus Dengue virus group
spellingShingle	Aedes aegypti Dengue RFLP RNA RT-PCR primer RNA Aedes aegypti amplicon Argentina article controlled study Dengue virus gene sequence mosquito nonhuman restriction fragment length polymorphism reverse transcription polymerase chain reaction RNA extraction RNA purification screening serotyping viral genetics virus detection Aedes Animals Argentina Base Sequence Dengue Dengue Virus Female Humans Insect Vectors Molecular Sequence Data Polymorphism, Restriction Fragment Length Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Nucleic Acid Aedes aegypti Dengue virus Dengue virus group Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
topic_facet	Aedes aegypti Dengue RFLP RNA RT-PCR primer RNA Aedes aegypti amplicon Argentina article controlled study Dengue virus gene sequence mosquito nonhuman restriction fragment length polymorphism reverse transcription polymerase chain reaction RNA extraction RNA purification screening serotyping viral genetics virus detection Aedes Animals Argentina Base Sequence Dengue Dengue Virus Female Humans Insect Vectors Molecular Sequence Data Polymorphism, Restriction Fragment Length Reverse Transcriptase Polymerase Chain Reaction Sequence Alignment Sequence Homology, Nucleic Acid Aedes aegypti Dengue virus Dengue virus group
description	Most molecular protocols for Dengue virus detection described so far are time consuming and cumbersome with mosquito samples. In order to count with a sensitive and specific molecular detection system for monitoring possible Dengue outbreaks and circulating viral serotypes in field-caught Aedes aegypti populations from Northeastern Argentina, a RT-PCR and RFLP assay was developed. The original RT-PCR assay proposed by Sudiro et al. for human serum was optimized for mosquito samples. Modifications were done at the RNA extraction-purification and at the thermal profile steps. The generic 230 bp amplicon was validated by RFLP assay and cycle sequencing. Results showed that, due to the generic characteristic of the primers used, certain mosquito genome regions could be co-amplified, making confirmation of the Dengue specific amplicon by RFLP assay a required step. Under these conditions, the proposed method can be employed as a Dengue viral generic screening procedure in Aedes aegypti mosquito samples, giving in our hands an estimated 99.52% of confirmed negatives (207/208 tested samples).
title	Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
title_short	Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
title_full	Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
title_fullStr	Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
title_full_unstemmed	Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina
title_sort	molecular detection of dengue viruses in field caught aedes aegypti mosquitoes from northeastern argentina
publishDate	2005
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01874640_v47_n3-4_p82_Liotta http://hdl.handle.net/20.500.12110/paper_01874640_v47_n3-4_p82_Liotta
_version_	1768544129292173312

Molecular detection of dengue viruses in field caught Aedes aegypti mosquitoes from northeastern Argentina

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