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One-step method for gametocyte and erythrocyte stage parasite detection published in malaria journal5/19/2017
The large collaborative team's work on spliced gametocyte mRNAs as infection detection markers is now published in the Malaria Journal (http://rdcu.be/sbZ4). Gametocytes are sexual-stage parasites that cause no disease in humans but are rather the forms transmitted from humans back to mosquitoes during a mosquito bite. The male and female gametocytes mate in the mosquito and allow the lifecycle to continue (see Lifecycle page). Since people have no symptoms from gametocytes and since most treatments for malaria infection do not kill gametocytes, these forms can circulate for a long time in humans and this can aid onward transmission to mosquitoes. Normal molecular diagnostic tests (like our 18S rRNA RT-PCR assay) cannot be used to detect gametocytes because all malaria parasites make 18S rRNAs and because other genomic targets are not stage-specific. Instead, the usual approach has been to perform gametocyte mRNA-specific RT-PCR AFTER treating extracted nucleic acids with DNase to destroy genomic material that would otherwise confuse the test. Most gametocyte-specific mRNAs in common use are from single exon genes. However, we (and a few others) have identified spliced gametocyte specific mRNAs and designed RT-PCR reagents that specifically amplify regions across these splicing sites, thereby eliminating the need for DNase treatment. The bottom line is that our new assay can be combined with the existing 18S rRNA RT-PCR assay in a ultrasensitive, multiplex assay that may be useful for field studies, drug studies and other clinical trials. We thank our colleagues at UW, the Center for Infectious Disease Research, Medicines for Malaria Venture, University of California San Francisco, Infectious Diseases Research Collaboration (Uganda), the London School of Hygiene & Tropical Medicine, PATH and the Seattle Malaria Clinical Trials Center (Fred Hutch Cancer Research Center) for collaborating on this project.
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