Identification of Vietnamese Coptotermes pest species based on the sequencing of two regions of 16S rRNA gene

Coptotermes, found in urban areas, is regarded as the most abundant building termite pest genus, widely distributed in Vietnam. The objectives of this study were to classify the Coptotermes found in certain provinces in Vietnam and assess the feasibility proposed PCR method by Szalanski et al., 2004 for identification of Coptotermes species. The proposed PCR method distinguishes species by the presence or absence of DNA fragments amplified with universal (LR-J-13007, LR-N-13398) and Coptotermes formosanus Shiraki specific (FST-F, FST-R) primer pairs. For this purpose, we collected six Coptotermes samples from five localities in Ha Noi (Van Quan-Ha Dong, Thai Ha-Dong Da, Vong Thi-Tay Ho, Tan Linh-Ba Vi, Bui Xuong Trach-Dong Da) and from Van Lam-Hung Yen province, Vietnam and employed the proposed simple PCR-based diagnosis for Coptotermes species (Szalanski et al., 2004). We amplified the universal 430-bp region from DNA templates of all six individual termite samples and the 151-bp regions of mtDNA 16S rRNA from DNA templates extracted from Tan Linh and Van Quan termites. Appearance of the species specific DNA fragments suggested that two Coptotermes samples can be classified as C. formosanus. However, by comparing the nucleotide sequences of the 430 bps and the 126 bps (eliminating the primer sequences from 151-bp region) with the corresponding sequences from Coptotermes species from GenBank, we found all six samples belonged to Coptotermes gestroi (Wasmann). Thus, in this case, PCR products were obtained employing primers with mismatches to the mtDNA 16S rRNA gene of C. gestroi. Therefore, the nucleotide sequencing is necessary for the identification of Coptotermes species including C. formosanus or C. gestroi based on the 430-bp and 151-bp regions of mtDNA 16S rRNA gene.