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Desarrollo y evaluación de una PCR multiplex realizada en un solo paso, para diferenciar estadíos acuáticos de la especie Aedes (Subgenus: Stegomyia) morfológicamente similares
Author(s) -
Das Biswadeep,
Swain Sunita,
Patra Aparna,
Das Mumani,
Tripathy Hari Kishen,
Mohapatra Namita,
Kar Santanu K.,
Hazra Rupenangshu K.
Publication year - 2012
Publication title -
tropical medicine and international health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.056
H-Index - 114
eISSN - 1365-3156
pISSN - 1360-2276
DOI - 10.1111/j.1365-3156.2011.02899.x
Subject(s) - biology , aedes , aedes aegypti , arbovirus , aedes albopictus , vector (molecular biology) , zoology , ecology , veterinary medicine , virology , larva , virus , genetics , medicine , recombinant dna , gene
Summary Objective To develop a single‐step multiplex PCR to differentiate the aquatic stages of Aedes aegypti , Aedes albopictus and Aedes vittatus collected from different breeding spots in arbovirus endemic/epidemic areas and to detect the most abundant species by the multiplex PCR. Methods Aquatic stages of different mosquito species were sampled by inspecting artificial and natural breeding sites in domestic and peridomestic areas. DNA was isolated from different stages of the three Aedes species. Using novel primers based on 18S rDNA sequence, a single‐step multiplex PCR was developed to clearly distinguish the three Aedes species. It was then evaluated in the aquatic stages of Aedes species collected from different areas. Results A total of 1150 aquatic stages were collected from 294 breeding spots, of which 156 contained Aedes species. Discarded tires were the major breeding spots of Aedes species. The aquatic stages were clustered into 230 pools; Ae. albopictus was detected in the largest number of pools, followed by Ae. aegypti and Ae. vittatus . Conclusions The Multiplex PCR clearly differentiated the aquatic stages of the three Aedes species and detected that Ae. albopictus was most profuse in different breeding spots surveyed, hence indicating to be the main vector in this region. So control measures can be designed against Ae. albopictus at an early stage to prevent any arboviral outbreak. This method is a convenient tool for precise identification of Aedes vectors during entomological surveys in arbovirus endemic/epidemic areas where several species coexist.