Molecular Detection of Plasmodium Falciparum from Malaria Diagnosed Patients attending Mater Hospital
Abstract
This research was aimed at evaluating the prevalence and molecular detection of Plasmodium falciparum from malaria diagnosed patients attending Mater Hospital. Venous blood samples (5ml) of 75 patients attending Mater Hospital were collected for preparation of thick blood firms for parasite screening and Nested PCR (nPCR) for Plasmodium falciparum (P. falciparum) gene detection. While thick film was prepared on microscopic slid, air dried, stained infield stain A and B.,and viewed under the light microscope with x100 objective lens, 10µl each of whole blood sample used for PCR Plasmodium falciparum gene detection using the nested Polymerase Chain Reaction (nPCR) as a diagnostic tool. The PCR products were analyzed in ethidium bromide stained 2% agarose gel. Microscopic examination of the stained blood film showed the presence of rig form trophozoites, schizonts and gametocytes of P. falciparum confirming high prevalence of malaria in Afikpo within the sampled population as 50 persons out of the 75 collected blood samples showed positive for malaria parasite. Age distribution of the samples shows that the most affected were those within the age brackets of 10 – 15 years with a prevalence rate of 26 (52%) followed by those within the age bracket of 16 – 21 years 11(22%). However, the nPCR analysis showed the presence of Plasmodium falciparum gene that resolved at 250bp and 270bp in all the 50 samples. Thereby indicating that Plasmodium falciparum was the prevalent specie that responsible for malaria in Afikpo. Following this result, it is recommended that PCR be included as part of the diagnostic tools for screening of the causative specie of malaria as this will go along way in ensuring effective treatment to prevent drug resistance.
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Introduction
In this part of the world, malaria has constituted itself the most fearful ailments all year round. Its prevalence has been such that no single day passes without a reference to it as amain cause of an illness. Its devastating health impact has been recognized to be transmitted by species of female anopheles mosquitoes. In 2003, Mbogo et al. opined that of the over 400 Anopheles species, only 30 – 40 can transmit malaria with Anopheles gambiae being the principal vector. Malaria is the world’smost deadly parasitic disease and is caused by infection with single-celled parasites of the genus Plasmodium belonging to the apicomplexan phylum. Anopheles mosquitoes transmit these parasites from one person to another in their bites (Microsoft Encarta, 2009). In Nigeria and rest of endemic Africa, the bulk of malaria episodes are attributable to P. falciparum. With an estimated 28 million cases and 38 000 deaths in 2011, malaria remains a significant public health problem in Sub-Saharan Africa (Olawole et al., 2014). The parasite destroys the red blood cells, leading to the clinical signs and symptoms such as fever, flu-like, chills, headache, muscle aches, tiredness, nausea, vomiting, diarrhea, and anemia and jaundice due to loss of red blood cells unless treated quickly the disease can kill within 24 hours: children under the age of five are particularly at risk (Wells et al., 2009). Malaria treatment has defied many known antimalarial drugs and so there is need to actually ascertain the exact parasite responsible for malaria transmission in Afikpo, Ebonyi State Niger. It is based on this that this study is on molecular detection of Plasmodium falciparum from malaria diagnosed patients attending mater hospital was conceived.
Conclusion
This study was aimed at Molecular detection of Plasmodium falciparum from malaria diagnosed patients attending Mater Hospital Afikpo in Ebonyi State. Out of the 75 samples analyzed 50 samples showed the presence of Plasmodium falciparum which was confirmed with application of Nested Polymerase Chain Reaction. The result indicated that P. falciparum was responsible for the transmission of malaria in Afikpo and therefore for adequate measures in preventing the spread of malaria in this area such as sleeping under mosquito nets and use of appropriate and efficient diagnostic tools such as PCR for precise identification of causative organisms so as to enable appropriate recommendation of antimalarial drugs to reduce or eliminate malaria resistance to drugs.
PCR should be used in confirming the actual causes of malaria illnesses so as to be specific in treatment: (i) People should adhere to the prescribed drugs for effective malaria treatment (ii) All breeding sites for mosquitoes should be eliminated so as to terminate the mosquitoes-malaria transmission.
