Nested multiplex PCR for identification and detection of human Plasmodium species including Plasmodium knowlesi
Journal Title: Asian Pacific Journal of Tropical Medicine - Year 2017, Vol 10, Issue 3
Abstract
Objective: To develop a new technique for diagnosis of Plasmodium knowlesi and at the same time to be able to discriminate among the diverse species of Plasmodium causing human malaria. Methods: In this study the nested multiplex malaria PCR was redesigned, targeting the 18S rRNA gene, to identify the fifth human Plasmodium species, Plasmodium knowlesi, together with the other human Plasmodium (Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale and Plasmodium malariae) by amplified fragment size using only two amplification processes and including an internal reaction control to avoid false negatives. Results: The technique was validated with 91 clinical samples obtained from patients with malaria compatible symptoms. The technique showed high sensitivity (100%) and specificity (96%) when it was compared to the reference method employed for malaria diagnosis in the Instituto de Salud Carlos Ⅲ and a published real-time PCR malaria assay. Conclusions: The technique designed is an economical, sensitive and specific alternative to current diagnosis methods. Furthermore, the method might be tested in knowlesimalaria endemic areas with a higher number of samples to confirm the quality of the method.
Authors and Affiliations
Maria Miguel-Oteo, Jos´e Miguel Rubio
Keywords
Related Articles
- EP ID EP278518
- DOI 10.1016/j.apjtm.2017.03.014
- Views 47
- Downloads 0
Antioxidants: Friend or foe?
Reactive oxygen species are the intermediates that are formed during the normal metabolic process which are effectively neutralized by the antioxidant system of the body. Any imbalance in this neutralization process caus...
Activation and IL-1b secretion of human peripheral phagocytes infected with Actinomadura madurae, Nocardia asteroides and Candida albicans
Objective: To evaluate the ability of Actinomadura madurae (A. madurae) and Nocardia asteroides (N. asteroides), using Candida albicans (C. albicans) as prototypic control, to elicit the activation and IL-1b secretion of...
Herbal remedies, vaccines and drugs for dengue fever: Emerging prevention and treatment strategies
Dengue fever is a disease that is caused by five dengue virus (DENV) serotypes. It is endemic in more than 128 countries. Millions of people are affected by this disease. But still, there is no specific treatment avail...
Potential of herbal constituents as new natural leads against helminthiasis: A neglected tropical disease
The WHO reports that billions of people and animals in tropical and subtropical regions are affected by helminthiasis as neglected tropical disease. It is predominant in underdeveloped areas; nevertheless, the increase i...
Inhibitory effects of methanolic Olea europaea and acetonic Acacia laeta on growth of Babesia and Theileria
Objective: To evaluate the antipiroplasmic activities of methanolic extract of Olea europaea (MOE) and acetonic extract of Acacia laeta (AAL) against Babesia and Theileria parasites in vitro and evaluate the chemotherape...