Molecular detection of Trichostrongylus species through PCR followed by high resolution melt analysis of ITS-2 rDNA sequences

被引：6
作者：
Arbabi, Mohsen [1 ,3 ]
Hooshyar, Hossein [1 ]
Lotfinia, Majid [2 ,3 ]
Bakhshi, Mohamad Ali [1 ]
机构：
[1] Kashan Univ Med Sci, Dept Med Parasitol, Kashan, Iran
[2] Kashan Univ Med Sci, Physiol Res Ctr, Basic Sci Res Inst, Kashan, Iran
[3] Kashan Univ Med Sci, Core Res Lab, Kashan, Iran
来源：
MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 2020年 / 236卷
关键词：
Trichostrongylus spp; Sheep; PCR; ITS-2-rDNA region; HRM analysis; Iran; INTERNAL TRANSCRIBED SPACER; RAPID DETECTION; RIBOSOMAL DNA; IDENTIFICATION; DIAGNOSIS; COLUBRIFORMIS; NEMATODA; HUMANS; INFECTIONS; PROVINCE;
D O I：
10.1016/j.molbiopara.2020.111260
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Polymerase chain reaction followed by high resolution melting (PCR-HRM) analysis is a simple, rapid and accurate method for molecular detection of various nematode species. The objective of the present study was, for the first time, to develop a PCR-HRM assay for the detection of various animal Trichostrongylus spp. A pair of primers targeting the ITS-2 rDNA region of the Trichostrongylus spp. was designed for the development of the HRM assay. DNA samples were extracted from 30 adult worms of Trichostrongylus spp., the ITS-2-rDNA region was amplified using PCR, and the resultant products were sequenced and characterized. Afterwards, the PCR-HRM analysis was conducted to detect and discriminate Trichostrongylus spp. Molecular sequence analysis revealed that 24, 4, and 1 of the samples were T. colubriformis, T. vitrinus and T. capricola, respectively. Results from PCR-HRM indicated that complete agreement was relatively found between speciation by HRM analysis and DNA sequencing for the detection of Trichostrongylus species. The PCR-HRM analysis method developed in the present study is fast and low-cost; the method can be comparable with other molecular detection techniques, representing a reliable tool for the identification of various species within the Trichostrongylus genus.
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页数：7
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