Malaria mitochondrial diagnosis: challenges and pitfalls
نویسندگان
چکیده
Background High-copy genomic sequences could be used as PCR targets for the detection of Plasmodium infections, providing increased sensitivity over single- or low-copy genes. Mitochondrial genomes malaria parasites are present in multiple copies a single mitochondrion, and each parasite has many mitochondria. Here, we describe development seven species-specific qPCR assays diagnosis vivax falciparum , targeting coding non-coding mitochondrial regions. Methods The optimization protocols involved gradient annealing temperatures concentrations primers probes, well inclusion additives/enhancers [e.g., dimethyl sulfoxide (DMSO), glycerol, bovine serum albumin (BSA)] to improve specificity amplification. Results Non-specific amplification other species human was observed different levels all assays. Regardless late Cq values most non-specific amplifications, application cutoff value did not completely exclude false-positive amplification, compromising also Conclusions Therefore, although have higher sensitivity, they frequently lose due their high sequence conservation. A screening evaluate cross-reaction between malaria-free samples must performed
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ژورنال
عنوان ژورنال: Frontiers in tropical diseases
سال: 2023
ISSN: ['2673-7515']
DOI: https://doi.org/10.3389/fitd.2023.1204195