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Original Article

Med Arch. 2021; 75(6): 418-423


Redesigning Primer of ITS2 (Internal Transcribed Spacer 2) for Specific Molecular Characterization of Malaria Vectors Anopheles Species

Kartika Senjarini, Muhammad Khalid Abdullah, Nuril Azizah, Miatin Alvin Septianasari, Ahmad Tosin, Rike Oktarianti, Syubbanul Wathon, Tri Yudani Mardining Raras.




Abstract

Background: Anopheles mosquitoes are vectors of malaria, which is a serious health issue in Indonesia. Thus, vector control is an important approach taken to overcome this disease. The first and most important step in vector control is vector identification. As some Anopheles species share similar morphological features, molecular identification helps make the process more accurate by using specific Objective: Our objective was to redesign a specific PCR primer for Anopheles species. Methods: The redesigned primer, named sma-ITS2, was then tested using mosquito samples from the Anopheles genus and other genera. Each mosquito was identified morphologically and their genomes were extracted. DNA samples were then amplified using the redesigned primer. Results: The sma-ITS2 primer pair was capable of amplifying ITS2 sequences from all of the Anopheles samples and unable to amplify any of the non-Anopheles samples, suggesting that it is specific to Anopheles only. Conclusion: DNA sequences as molecular markers. Internal Transcribed Spacer 2 (ITS2) is a non-coding DNA region commonly used as a molecular marker for the DNA barcoding of insects. Many of the available ITS2 primers are universally designed for insects and, as such, are typically less specific for identifying certain genera, such as Anopheles sp. Therefore, redesigning a specific ITS2 primer is needed for specific Anopheles identification. The sma-ITS2 primer pair was able to identify intra-species of Anopheles, but its efficiency in making differentiations within a species complex should be evaluated further.

Key words: Primer, ITS2, Anopheles, Vector, Malaria






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