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Developing molecular diagnostics for detection of red imported fire ants using two genes, Sinv11108 and Sinv11977
Author(s) -
Kim JiHye,
Park Youngjin,
Kim Woo Jin,
Kim AYoung,
Nguyen Phuong,
Lyu Dong Pyo,
Lee Heung Sik,
Koh Young Ho
Publication year - 2019
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21610
Subject(s) - biology , agarose gel electrophoresis , polymerase chain reaction , agarose , taqman , multiplex polymerase chain reaction , gene , gel electrophoresis , dna , molecular diagnostics , microbiology and biotechnology , computational biology , dna extraction , genetics
Aggressive red imported fire ants (RIFAs) are expanding their habitat due to active international trade and global warming. To prevent infestation and settlement, RIFAs must be removed during the quarantine process. Because RIFAs are social insects and have different morphological characteristics depending on their castes, non‐ant taxonomists have difficulty confirming RIFAs based on their morphological characteristics alone. The disadvantages of previously reported RIFA molecular diagnostics are that they require additional steps, such as restriction enzyme digestion followed by agarose gel electrophoresis separation or DNA sequence verification for polymerase chain reaction (PCR)‐amplified products. To overcome these drawbacks, two RIFA‐specific genes were selected and used to develop diverse PCR‐based RIFA molecular diagnostic techniques. We found that RIFAs could be confirmed by conventional PCR targeting of two RIFA‐specific genes followed by agarose electrophoresis separation. In addition, TaqMan probe real‐time PCR methods had the advantage of confirming RIFAs immediately after the reactions were completed by observing fluorescence indexes. Finally, multiplex PCRs enhanced RIFA specificity and sensitivity. The new molecular diagnostic methods developed in this study had the advantages of reducing false positive and negative results together with high specificity and sensitivity for RIFAs.