Allele Specific Polymerase Chain Reaction Kullanilmasi

• Published in: Türkiye parazitolojii dergisi 2022-12, Vol.46 (4), p.307
• Main Authors: Yavasoglu, Sare Ilknur, Simsek, Fatih Mehmet
• Format: Article
• Language: English
• Subjects: Anopheles; Genetic aspects; Malaria; Polymerase chain reaction; Population genetics
• ISSN: 1300-6320
• DOI: 10.4274/tpd.galenos.2022.20592

Objective: The study aims to determine the presence of L1014F, L1014S, L1014C alleles, which are responsible for knockdown resistance and Ace-1 G119S alleles, which are responsible for acetylcholinesterase insensitivity in Anopheles superpictus, the secondary vector of malaria in Turkey. Methods: In this study, 60 Anopheles superpictus adult females were collected from Aydin, Denizli, and Mugla provinces. Then, allele-specific primers for kdr L1014F, L1014S, and L1014C alleles, and the Ace-1 G119S allele were designed. The presence of these alleles was screened in three Anopheles superpictus populations by allele-specific polymerase chain reaction. Results: Although L1014S allele frequency was too low in Aydin, Mugla, and Denizli populations, neither kdr L1014F and L1014C nor Ace-1 G119S mutations were found in any population. Conclusion: In this study, kdr L1014S mutation was detected for the first time in the Aegean Anopheles superpictus populations. Keywords: Anopheles superpictus, knockdown, acetylcholinesterase, allele-specific primer Amaç: Bu çalismanin amaci, Türkiye'de sitmanin ikincil vektörü olan Anopheles superpictus'ta knockdown direncinden sorumlu olan L1014F, L1014S, L1014C allellerinin ve asetilkolinesteraz duyarsizligindan sorumlu olan Ace-1 G119S allellerinin varligini tespit etmektir. Yöntemler: Aydin, Denizli ve Mugla illerinden 20'ser adet Anopheles superpictus ergin disileri toplanmistir. Daha sonra knockdown direncinden sorumlu olan L1014F, L1014S ve L1014C allellerine ve Ace-1 G119S alleline özgü primerler tasarlanarak allele özgü polimeraz zincir reaksiyonu ile bu üç Anopheles superpictus popülasyonunda bu allellerin varligi taranmistir. Bulgular: Aydin, Mugla ve Denizli popülasyonlarinda çok düsük oranda L1014S mutasyonuna rastlanmakla birlikte, popülasyonlarin hiçbirinde ne kdr L1014F ve L1014C; ne de Ace-1 G119S mutasyonuna rastlanmistir. Sonuç: Bu çalismanin sonucu olarak, ilk kez Anopheles superpictus Ege bölgesi popülasyonlarinda kdr L1014S mutasyonu tespit edilmistir. Anahtar Kelimeler: Anopheles superpictus, knockdown, asetilkolinesteraz, allele özgü primer