Establishment of a multiplex PCR method for the identification of Anopheles splendidus,Anopheles maculatus and Anopheles aconitus
NIE Guoai
ZHANG Rong
CAI Liping
SU Xiaojian
YANG Guoping
GENG Heyuan
HE Ji
TAN Weilong
YANG Qinggui
Abstract：Objective To establish a multiple polymerase chain reaction(PCR)method for simultaneous,accurate,and rapid identification of Anopheles splendidus,An.maculatus and An.aconitus.Methods Specific primers targeting the mitochondrial COI gene of An.splendidus,An.maculatus and An.aconitus were designed.The PCR conditions were optimized,and the reliability of the assay was evaluated through specificity and sensitivity tests.Results The primers for An.splendidus,An.maculatus and An.aconitus exhibited identical optimal concentrations and demonstrated high specificity at an annealing temperature of 58℃,amplifying distinct fragments of 66,135,and 299 bp,respectively.No amplification was observed for non-target species or blank controls.The detection limit for all three species was determined to be 10-4 ng/μL.Conclusion This multiplex PCR method enables simultaneous,precise,and rapid identification of An.splendidus,An.maculatus and An.aconitus,even in damaged or non-adult specimens where morphological features are obscured.This molecular tool provides a valuable resource for monitoring mosquito population dynamics and predicting arthropod-borne diseases risks.And it is of practical significance for improving the precise vector control systems.
Keywords：Anopheles splendidusAnopheles maculatusAnopheles aconitusmultiplex PCR
Publication Date：2025-12-20
Online Publishing Date：2026-01-09(First online date of this platform, not the publication date of the document)
Pages：5( 679-683 )