Semi-quantitative differentiation of cyathostomin larval cultures by reverse line blot

Veterinary Parasitology
F N J KooymanJ A Wagenaar

Abstract

Cyathostomins are the most prevalent horse nematodes worldwide and over 50 species are described. The eggs and the infective larvae (L3) can easily be obtained or cultured from infected horses, but cannot be differentiated morphologically at species level. A reverse line blot (RLB) method based on the hybridization of a PCR fragment with a species specific probe, has previously been developed for the differentiation of individual eggs and/or L3s, but is too labor intensive for large scale studies. In the present study a RLB method on multiple pooled L3s for the semi-quantitative differentiation of cyathostomin larval cultures was developed and validated. First, the probability of the presence of a certain species within a pool was calculated as function of the frequency and the number of L3s within a pool. Ten L3s per pool were found to be optimal. Next, the probability, the chance of occurrence was calculated when 4 pools per culture were used. The probability distributions for 0, 1, 2, 3 or 4 positive pools were transformed into the corresponding median frequency of the cumulative probability: 0.014, 0.04, 0.08, 0.16 and 0.59, respectively. Based on these calculated probabilities, RLB on 10 L3s per pool and 4 pools per sample...Continue Reading

References

Apr 30, 1999·International Journal for Parasitology·G C HungR B Gasser
Mar 10, 2001·International Journal for Parasitology·J E HodgkinsonJ B Matthews
Nov 26, 2002·Veterinary Parasitology·C Collobert-LaugierP Dorchies
Sep 21, 2010·Veterinary Parasitology·D C K van DoornH W Ploeger
Jan 16, 2013·Veterinary Parasitology·Ricardo J CaneverMarcelo B Molento

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Citations

Jan 27, 2018·Parasites & Vectors·Rebecca A MolenaCinzia Cantacessi
Jul 14, 2017·Frontiers in Cellular and Infection Microbiology·Christina M BredtmannGeorg von Samson-Himmelstjerna
Jun 3, 2021·Animals : an Open Access Journal From MDPI·Alexa C B Johnson, Amy S Biddle

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