Development of cytochrome c oxidase subunit 1 gene-based duplex PCR for identifying Clonorchis sinensis and Haplorchis taichui

Abstract

Abstract:

Genomic DNA was extracted from Clonorchis sinensis and Haplorchis taichui, primers for cytochrome c oxidase subunit 1(COX1) gene were designed, respectively, and single and duplex PCR were performed. Single PCR for C. sinensis using three pairs of primers, FC1/RC1 and FC3/RC3 primer pairs generated specific bands of 328 bp and 356 bp, respectively, while no band was produced when using the FC2/RC2 primer pair. Similarly, single PCR using FH1/RH1 and FH2/RH2 primer pairs for H. taichui produced specific bands of 200 bp and 190 bp, respectively. Furthermore, duplex PCR combining the FC1/RC1 primer pair with FH1/RH1 or FH2/RH2 primer pair resulted in specific bands for both C. sinensis and H. taichui, with no cross-species reaction.

Key words: Clonorchis sinensis, Haplorchis taichui, Cytochrome c oxidase subunit 1 gene, Duplex PCR

CLC Number:

R383.2

Zhi-hua JIANG, Qing-li YANG, Yi-chao YANG. Development of cytochrome c oxidase subunit 1 gene-based duplex PCR for identifying Clonorchis sinensis and Haplorchis taichui[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2018, 36(2): 187-189.

Figures/Tables 4

References 14

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名称	GenBank登录号	引物名称	序列5′→3′	位置/nt	扩增条带长度/bp
华支睾吸虫	AF096229^a	FC1	GCCTATAGTGAAAAGCACCA	104~431	328
		RC1	GTTAATATTGCCGGGGTTTG
		FC2	ACAAAACCCGGATTATCCAA	124~449	326
		RC2	GCATCCTGAGGTTTATGTGT
		FC3	TAAACCCCGCCTATAGTGAA	96~451	356
		RC3	GGGCATCCTGAGGTTTATG
扇棘单睾吸虫	EF055885^b	FH1	TGGGTTGGATGTTAAGACG	154~353	200
		RH1	AAGAAGACAACACAATCCCG
		FH2	AGACACATTTGTAGGACGTTA	32~221	190
		RH2	TACCTGTGGGGACTCCTA

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