Abstract:
:The purpose of this study was to evaluate the practicality of using real-time PCR for quantifying feces-derived trichostrongyle eggs. Haemonchus contortus eggs were used to evaluate fecal contaminants, time after egg embryonation, and the presence of competing and non-competing DNAs as factors that might interfere with generating reproducible results during simplex and multiplex quantitative real-time PCR (QPCR). Real-time PCR results showed linear quantifiable amplification with DNA from five to 75 eggs. However, threshold cycle (C (T)) values obtained by amplification of DNA from egg numbers between 75 and 1,000 did not differ significantly. Inhibitors of QPCR were effectively removed during DNA extraction as exemplified by the absence of any improvement in C (T) values with bovine serum albumin or phytase treatments. Changes from egg embryonation could only be detected during the first 6 h. Noncompetitive DNA did not appear to impact amplification; however, in a multiplex reaction a competing trichostrongyle such as Cooperia oncophora can hinder amplification of H. contortus DNA, when present at tenfold greater amounts. This study demonstrates the usefulness of QPCR for amplification and quantification of trichostrongyle eggs, and identifies potential limitations, which may be addressed through multiplex assays or the addition of a standard: exogenous DNA target.
journal_name
Parasitol Resjournal_title
Parasitology researchauthors
Harmon AF,Williams ZB,Zarlenga DS,Hildreth MBdoi
10.1007/s00436-006-0428-0subject
Has Abstractpub_date
2007-06-01 00:00:00pages
71-6issue
1eissn
0932-0113issn
1432-1955journal_volume
101pub_type
杂志文章abstract::Praziquantel is still very effective for the treatment of schistosomiasis, but there are rising concerns on the potential risk of developing resistances because of the extensive use of this drug. Triclabendazole, a systemic anthelmintic, is very effective against other trematodes such as Paragonimus spp. and Fasciola ...
journal_title:Parasitology research
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journal_title:Parasitology research
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pub_type: 杂志文章
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journal_title:Parasitology research
pub_type: 杂志文章,评审
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journal_title:Parasitology research
pub_type: 杂志文章
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pub_type: 杂志文章,评审
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journal_title:Parasitology research
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