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PHD : Parasites, Hosts and Diseases

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Brief Communication

Differential diagnosis of Trichostrongylus and hookworm eggs via PCR using ITS-1 sequence

The Korean Journal of Parasitology 2007;45(1):69-74.
Published online: March 20, 2007

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Corresponding author (tsyong212@yumc.yonsei.ac.kr)
• Received: October 10, 2006   • Accepted: January 5, 2006

Copyright © 2007 by The Korean Society for Parasitology

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Differential diagnosis of Trichostrongylus and hookworm eggs via PCR using ITS-1 sequence
Korean J Parasitol. 2007;45(1):69-74.   Published online March 20, 2007
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Differential diagnosis of Trichostrongylus and hookworm eggs via PCR using ITS-1 sequence
Image Image Image Image Image
Fig. 1 Trichostrongylus colubriformis adults collected from humans in Khamouane Province, Laos in 2003. (A) A female worm showing uterine eggs in one row. (B) Spicules of a male worm.
Fig. 2 Phylogenetic relationship of the ITS regions from Trichostrongylus spp by using the PAUP* (version 4.0) program. The data were retrieved from GenBank except* T. colubriformis, of which was obtained in this study.
Fig. 3 Nucleotide sequence alignment of the ITS-1 gene from Trichostrongylus colubriformis used in this study, Ancylostoma duodenale, and Necator americanus. The locations of the forward primers jmAD (5'-TGCGAAGTTCGCGTTTCGCTGAGCT-3'), jmNA (5'-CGTTAACATTGTATACCTGTACATAC-3'), and jhTsp (5'-TTATGTGCCACAAATGAAGA-3') are underlined in each line. Asterisks indicate identical nucleotides.
Fig. 4 Schematic representation of part of the rDNA transcriptional unit and relative locations of primers (NC5, NC2, jmAD, jmNA and jhTsp) used in this study. The region harboring ITS 1, ITS 2, and 5.8 S rDNA was amplified with NC5 and NC2 primers. The species-specific primers, jmAD, jmNA, and jhTsp were targeted to the region within the ITS-1 sequence.
Fig. 5 Specificity of ITS-PCR in the recognition of the genomic DNA of other bacteria, protozoa, Lane M, 100 bp ladder, 600 bp and 100 bp bands were shown. Lane 1; N. americanus, Lane 2; A. duodenale, Lane 3; Trichostrongylus colubriformis, Lane 4; Escherichia coli, Lane 5; Entamoeba histolytica, Lane 6; Giardia lamblia. Only specifically amplified PCR products were detected on lanes 1-3.
Differential diagnosis of Trichostrongylus and hookworm eggs via PCR using ITS-1 sequence