ArticlePathogens (Basel, Switzerland)2021
Differing Effects of Standard and Harsh Nucleic Acid Extraction Procedures on Diagnostic Helminth Real-Time PCRs Applied to Human Stool Samples.
Article in Pathogens (Basel, Switzerland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Optimisation and analytical assessment of a TaqMan™ probe-based real-time PCR assay designed to diagnose infection with Schistosoma japonicum.Parasites & vectors · 2026Article
- Matrix-Dependent Sensitivity of Two Pan-Trematode PCR Assays for DetectingInfectious disease reports · 2026Article
- Intestinal Helminth Infections in Ghanaian Children from the Ashanti Region between 2007 and 2008-A Retrospective Cross-Sectional Real-Time PCR-Based Assessment.Tropical medicine and infectious disease · 2022Article
- Enteric Bacteria and Parasites with Pathogenic Potential in Individuals of the Colombian Indigenous Tribe Kogui.Microorganisms · 2022Article
- Comparison of the Influence of Different Nucleic Acid Extraction Assays on the Sensitivity ofMicroorganisms · 2022Article
- Evaluation of Commercial Concentration Methods for Microscopic Diagnosis of Protozoa and Helminths in Human Stool Samples in a Non-Endemic Area.Microorganisms · 2022Article
- Multicentric Evaluation of SeeGene Allplex Real-Time PCR Assays Targeting 28 Bacterial, Microsporidal and Parasitic Nucleic Acid Sequences in Human Stool Samples.Diagnostics (Basel, Switzerland) · 2022Article
- Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis ofPathogens (Basel, Switzerland) · 2021Article
- Low Sensitivity of Real Time PCRs Targeting Retrotransposon Sequences for the Detection ofPathogens (Basel, Switzerland) · 2021Article
- New Developments in PCR-Based Diagnostics for Bacterial Pathogens Causing Gastrointestinal Infections-A Narrative Mini-Review on Challenges in the Tropics.Tropical medicine and infectious disease · 2021Review
- Evaluation of the Allplex™ GI-Helminth(I) Assay, the first marketed multiplex PCR for helminth diagnosis.Parasite (Paris, France) · 2021Article
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11 authors.
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Abstract
This study aimed to assess standard and harsher nucleic acid extraction schemes for diagnostic helminth real-time PCR approaches from stool samples. A standard procedure for nucleic acid extraction from stool and a procedure including bead-beating as well as proteinase K digestion were compared with group-, genus-, and species-specific real-time PCR assays targeting helminths and nonhelminth pathogens in human stool samples. From 25 different in-house and commercial helminth real-time PCR assays applied to 77 stool samples comprising 67 historic samples and 10 external quality assessment scheme samples positively tested for helminths, higher numbers of positive test results were observed after bead-beating-based nucleic acid extraction for 5/25 (20%) real-time PCR assays irrespective of specificity issues. Lower cycle threshold values were observed for one real-time PCR assay after the standard extraction scheme, and for four assays after the bead-beating-based scheme. Agreement between real-time PCR results after both nucleic acid extraction strategies according to Cohen's kappa ranged from poor to almost perfect for the different assays. Varying agreement was observed in eight nonhelminth real-time PCR assays applied to 67 historic stool samples. The study indicates highly variable effects of harsh nucleic acid extraction approaches depending on the real-time PCR assay used.
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