Quantitative PCR for Leishmania infantum DNA Detection
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Questions and Answers

The article 'Comparison of Four Diagnostic Methods for Detection and Relative Quantification of Haemonchus contortus Eggs in Feces Samples' was published in 2016.

False

The ISBN of the book 'Advances in Parasitology' is 978-0-12-810395-1.

True

The study 'Nematode Parasitism Affects Lying Time and Overall Activity Patterns in Lambs Following Pasture Exposure around Weaning' was published in 2020.

False

The authors of the article 'Molecular Detection of Two Major Gastrointestinal Parasite Genera in Cattle Using a Novel Droplet Digital PCR Approach' are Baltrušis, P., Halvarsson, P., and Höglund, J.

<p>True</p> Signup and view all the answers

The article 'Egg Production Dynamics and Fecundity of Heterakis gallinarum Residing in Different Caecal Environments of Chickens Induced by Fibre-Rich Diets' was published in 2013.

<p>False</p> Signup and view all the answers

Study Notes

qPCR vs ddPCR

  • qPCR does not provide absolute quantification and lacks a standardized method.
  • ddPCR is advantageous over qPCR as it does not require a standard curve and is easy to standardize.
  • A ddPCR assay for Leishmania infantum DNA detection was developed and validated, showing perfect agreement with qPCR.

Theileria spp.

  • Diagnosis of bovine theileriosis is based on clinical signs and detecting parasites in blood samples with light microscopy.
  • Species identification is pivotal in bovine theileriosis.

Haemonchus contortus

  • A ddPCR assay targeted dyf-7 SNPs as a molecular marker for ivermectin resistance in H. contortus strains.
  • The assay was robust for detecting mutations of the dyf-7 allele frequency in mixed larval cultures with a low threshold (≈3 copies/µL).
  • However, the study showed that mutations in dyf-7 are not involved in ivermectin resistance.

Mixed Infections in Ruminants

  • Mixed infections with gastrointestinal nematodes are especially important in cases of low levels of H. contortus infection that are difficult to detect with conventional parasitological techniques.
  • A ddPCR assay was developed and validated for the absolute quantification and identification of three genera of strongylids in sheep: Haemonchus, Teladorsagia, and Trichostrongylus.
  • The assay employed one universal target for all strongylid gastrointestinal nematodes and three genus-specific targets, allowing the estimation of the relative abundance of each parasite species with high precision.
  • The ddPCR assay presented a good agreement with FECRT and no cross-amplifications were observed between the genera.

Dual Infections in Poultry

  • The flotation technique is sensitive but difficult to differentiate Ascaridia galli and Heterakis gallinarum due to morphological similarities.
  • A duplex ddPCR was developed to identify and quantify A. galli and H. gallinarum DNA in chicken feces, showing substantial agreement with the flotation technique.
  • Early detection and treatment of H. gallinarum infection are crucial in flocks with a history of histomonosis.

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Description

This quiz assesses your understanding of quantitative PCR (qPCR) and droplet digital PCR (ddPCR) methods for detecting Leishmania infantum DNA, including their limitations and applications.

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