DUGi

Aplicació de la PCR digital pel diagnòstic de la malària

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Malaria or paludism is a vector infection caused by parasites of the genus Plasmodium and transmitted by female mosquitoes of the genus Anopheles. This disease is a public health problem, especially in Africa, where 90% of cases are concentrated. The initial symptoms are non-specific, which, if not treated in time, can be complicated and fatal. There are effective drugs for treatment, but resistance to them is increasingly common. The diagnosis of malaria is carried out using microscopic, immunological and/or molecular methods, with the amplification of the parasite genetic material being the principle of molecular methods. The most used molecular technique is PCR, but new promising methods have recently appeared, such as digital PCR (dPCR). The objective of this TFG was to study the dPCR and its application in diagnosing malaria to evaluate it as a possible diagnostic alternative. The bibliographic search was done from the Web of Science database by combination of the following keywords: "malaria", "Plasmodium", "digital PCR", "dPCR" and "ddPCR". Finally, 12 articles were selected after reviewing the titles and abstracts results. dPCR is an innovative molecular tool that allows absolute quantification by partitioning the reaction. A variant of the dPCR is the droplet digital PCR (ddPCR), which shares the same principle as the dPCR but differs in the partitioning of the sample. Whole blood is the most suitable sample for diagnosis, but serum, saliva, or oral swabs may be viable alternatives. The main advantages of dPCR include that it does not require a standard curve for quantification, high tolerance to inhibitors, highly sensitive, very useful for target DNA detection in samples with low parasitic load and mixed infections, and the ability to detect rare genetic variants with great precision. However, the dPCR has some weaknesses, such as the possible saturation of the reaction in very concentrated samples and being expensive and laborious technique. For the future, dPCR is a promising technique for the diagnosis of malaria, but its application in endemic areas with limited resources requires the development of more affordable equipment. The main applications of the dPCR include diagnosis, analysis of drug resistance against the parasite, and vector control of the disease

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