ArticleFrontiers in microbiology2015
Quantitative RT-PCR assay for high-throughput screening (HTS) of drugs against the growth of Cryptosporidium parvum in vitro.
Article in Frontiers in microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- A rapid sporozoite viability assay identifies leads and targetable enzymatic activities against the zoonotic Cryptosporidium parvum.International journal for parasitology. Drugs and drug resistance · 2026Article
- 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
- Pathogenicity and virulence ofVirulence · 2025Review
- Targeting translation initiation yields fast-killing therapeutics against the zoonotic parasite Cryptosporidium parvum.PLoS pathogens · 2025Article
- High-Throughput Fluorescence Screening Enables Globally Consistent Identification of ABA Signaling Modulators.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Rapid review: Recent advances inCurrent research in parasitology & vector-borne diseases · 2025Review
- Cryptosporidium uses CSpV1 to activate host type I interferon and attenuate antiparasitic defenses.Nature communications · 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
- Octaarginine Improves the Efficacy of Nitazoxanide againstPathogens (Basel, Switzerland) · 2022Article
- The mucin-like, secretory type-I transmembrane glycoprotein GP900 in the apicomplexan Cryptosporidium parvum is cleaved in the secretory pathway and likely plays a lubrication role.Parasites & vectors · 2022Article
- Comparative Characterization of CpCDPK1 and CpCDPK9, Two Potential Drug Targets Against Cryptosporidiosis.Microorganisms · 2022Article
- A high-throughput screening RT-qPCR assay for quantifying surrogate markers of immunity from PBMCs.Frontiers in immunology · 2022Article
- Implication of Potential Differential Roles of the Two Phosphoglucomutase Isoforms in the Protozoan ParasitePathogens (Basel, Switzerland) · 2021Article
- Unique Tubulin-Based Structures in the Zoonotic Apicomplexan ParasiteMicroorganisms · 2021Article
- A Single-Pass Type I Membrane Protein from the Apicomplexan ParasiteMicroorganisms · 2021Article
- Discovery of New Microneme Proteins inFrontiers in veterinary science · 2021Article
- Novel Antiparasitic Activity of the Antifungal Lead Occidiofungin.Antimicrobial agents and chemotherapy · 2020Article
- Cryptosporidium parvum Elongation Factor 1α Participates in the Formation of Base Structure at the Infection Site During Invasion.The Journal of infectious diseases · 2020Article
- Characterization of Three Calcium-Dependent Protein Kinases ofFrontiers in microbiology · 2020Article
- 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
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2 authors.
Funding
Abstract
Our laboratory has previously developed a qRT-PCR assay to assess drug efficacy on the growth of Cryptosporidium parvum in vitro by detecting the levels of parasite 18S rRNA. This approach displayed up to four orders of magnitude of linear dynamic range and was much less labor-intensive than the traditional microscopic methods. However, conventional qRT-PCR protocol is not very amendable to high-throughput analysis when total RNA needs to be purified by lengthy, multi-step procedures. Recently, several commercial reagents are available for preparing cell lysates that could be directly used in downstream qRT-PCR analysis (e.g., Ambion Cell-to-cDNA kit and Bio-Rad iScript sample preparation reagent). Using these reagents, we are able to adapt the qRT-PCR assay into high-throughput screening of drugs in vitro (i.e., 96-well and 384-well formats for the cultivation of parasites and qRT-PCR detection, respectively). This qRT-PCR protocol is able to give a >150-fold linear dynamic range using samples isolated from cells infected with various numbers of parasites. The new assay is also validated by the NIH-recommended intra-plate, inter-plate, and inter-day uniformity tests. The robustness and effectiveness of the assay are also confirmed by evaluating the anti-cryptosporidial efficacy of paromomycin and by a small scale screening of compounds.
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