ArticleAntimicrobial agents and chemotherapy2005
Application of quantitative real-time reverse transcription-PCR in assessing drug efficacy against the intracellular pathogen Cryptosporidium parvum in vitro.
Article in Antimicrobial agents and chemotherapy, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers.
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Who cites it
55 citing papers in PubMed, 80 citations in OpenAlex.
- Phenotypic High-Throughput Screening and Drug Efficacy Test Against the Growth of Cryptosporidium parvum In Vitro by qRT-PCR.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Single Oocyst Selection and Whole-Genome Amplification of Cryptosporidium parvum.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Host cell KRT-18 as a non-redox dependent interactor of Cryptosporidium parvum PDI enables parasite infection.Communications biology · 2025Article
- Article
- A single-pass type I membrane protein, mannose-specific L-type lectin, potentially involved in the adhesion and invasion of Cryptosporidium parvum.Parasite (Paris, France) · 2024Article
- Functional characterization of CpADF, an actin depolymerizing factor protein in Cryptosporidium parvum.Parasitology research · 2023Article
- On-target inhibition of Cryptosporidium parvum by nitazoxanide (NTZ) and paclitaxel (PTX) validated using a novel MDR1-transgenic host cell model and algorithms to quantify the effect on the parasite target.PLoS neglected tropical diseases · 2023Article
- Article
- Comparative Characterization of CpCDPK1 and CpCDPK9, Two Potential Drug Targets Against Cryptosporidiosis.Microorganisms · 2022Article
- Characterization of Calcium-Dependent Protein Kinase 2A, a Potential Drug Target Against Cryptosporidiosis.Frontiers in microbiology · 2022Article
- The Oesophageal Squamous Cell Carcinoma Cell Line COLO-680N Fails to Support SustainedPathogens (Basel, Switzerland) · 2021Article
- Bicyclic azetidines kill the diarrheal pathogenScience translational medicine · 2020Article
- Potential therapeutic and prophylactic effects of Asafoetida in murine cryptosporidiosis.Journal of parasitic diseases : official organ of the Indian Society for Parasitology · 2020Article
- Article
- Characterization of Calcium-Dependent Protein Kinases 3, a Protein Involved in Growth ofFrontiers in microbiology · 2020Article
- Characterization of Three Calcium-Dependent Protein Kinases ofFrontiers in microbiology · 2020Article
- Identification of a potent benzoxaborole drug candidate for treating cryptosporidiosis.Nature communications · 2019Article
- The Action of the Hexokinase Inhibitor 2-deoxy-d-glucose on Cryptosporidium parvum and the Discovery of Activities against the Parasite Hexokinase from Marketed Drugs.The Journal of eukaryotic microbiology · 2019Article
- A suite of phenotypic assays to ensure pipeline diversity when prioritizing drug-like Cryptosporidium growth inhibitors.Nature communications · 2019Article
- The effect of autophagy on the survival and invasive activity of Eimeria tenella sporozoites.Scientific reports · 2019Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
We report here on a quantitative real-time reverse transcription-PCR (qRT-PCR) assay for assessing drug efficacy against the intracellular pathogen Cryptosporidium parvum. The qRT-PCR assay detects 18S rRNA transcripts from both parasites, that is, the cycle threshold for 18S rRNA from parasites (C(T)([P18S])) and host cells (C(T)([H18S])), and evaluates the relative expression between parasite and host rRNA levels (i.e., deltaC(T) = C(T)([P18S]) - C(T)([H18S])) to minimize experimental and operational errors. The choice of qRT-PCR over quantitative PCR (qPCR) in this study is based on the observations that (i) the relationship between the logarithm of infected parasites (log[P]) and the normalized relative level of rRNA (deltadeltaC(T)) is linear, with a fourfold dynamic range, by qRT-PCR but sigmoidal (nonlinear) by qPCR; and (ii) the level of RNA represents that of live parasites better than that of DNA, because the decay of RNA (99% in approximately 3 h) in dead parasites is faster than that of DNA (99% in approximately 24 to 48 h) under in vitro conditions. The reliability of the qRT-PCR method was validated by testing the efficacies of nitazoxanide and paromomycin on the development of two strains of C. parvum (IOWA and KSU-1) in HCT-8 cells in vitro. Both compounds displayed dose-dependent inhibitions. The observed MIC50 values for nitazoxanide and paromomycin were 0.30 to 0.45 micro/ml and 89.7 to 119.0 microg/ml, respectively, comparable to the values reported previously. Using the qRT-PCR assay, we have also observed that pyrazole could inhibit C. parvum development in vitro (MIC50 = 15.8 mM), suggesting that the recently discovered Cryptosporidium alcohol dehydrogenases may be explored as new drug targets.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.