KARAKTERISASI PLASMODIUM SPESIES MENGGUNAKAN METODE KONVENSIONAL DAN NESTED PCR DI KABUPATEN NIAS SELATAN PROPINSI SUMATERA UTARA INDONESIA

Harurikson, Lumbantobing (2017) KARAKTERISASI PLASMODIUM SPESIES MENGGUNAKAN METODE KONVENSIONAL DAN NESTED PCR DI KABUPATEN NIAS SELATAN PROPINSI SUMATERA UTARA INDONESIA. Doctoral thesis, Universitas Andalas.

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Abstract

Malaria is one of the most contagious diseases that are still a public health problem in the world including Indonesia. World Malaria Report 2011 mentions that malaria occurs in 106 countries, even 3.3 billion people live in areas at risk of contracting malaria. The number of malaria cases in the world is 216 million cases, of which 28 million cases occur in ASEAN. Each year as many as 660 thousand people died of malaria, especially toddlers (86%), 320 thousand of which are in Southeast Asia including Indonesia. Malaria is a parasitic infectious disease with the highest morbidity and mortality rates in North Sumatra. Particularly in South Nias district, there are two types of malaria: malaria vivax endemic and malaria falciparum. The diagnosis of malaria based on the discovery of parasites in thick and thin blood dams is still a problem in endemic areas due to equipment limitations and skill-dependent skills so as to diagnose newly forgotten species parasites so that rapid diagnosis and therapy are fundamental to stopping the transmission of malaria. In contrast, the Nested-2 PCR Biomolecular test can detect small amounts of parasites and new species plasmodium. SD Bioline Malaria P.f / Pan antigen consists of a strip membrane, which is re-coated with monoclonal antibody and 1 polyclonal antibody in the form of two separate lines on the surface of the test kit. The first monoclonal antibody (P.f test line) specific to HRP2 P.falciparum and the second polyclonal antibody (P.Pan test line) for lactate dehydrogenase Plasmodium falciparum and Plasmodium vivax) adsorbed to human lactate dehydrogenase. Bi SDH Bioline Malaria P.f / Pan is designed to diagnose Plasmodium falciparum and other Plasmodium species separately. Microscopy is a gold standard (standard ) for the exact diagnosis of malaria. Microscopic examination is done by making blood thick and thin. Blood and thin blood sample (SD) examination at hospital / Puskesmas / field to determine: The presence or absence of malaria parasites (positive or negative); Species and stages of Plasmodium; Parasite density. The purpose of this study was to investigate the sensitivity and specificity of diagnostic tests of P. falciparum, P. vivax, P.malariae, P.ovale, P.knowlesi infection in malaria endemic areas. So from the results of this study can be a contribution in the handling of cases of severe malaria and malaria of the brain for early detection in diagnosing malaria quickly and accurately so that treatment as soon as possible. The research was conducted at the Biomolecular Laboratory Faculty of Medicine, Prima University, Medan. The research sample was obtained from Puskesmas Pulau Tello Kecamatan Pulau-Pulau Batu, South Nias District, from November 2014 until November 2016. The study population was malaria patients who visited Toso Island Community Health Center, Pulau Batu Island, South Nias District. Patients with a diagnosis of Clinical Malaria and Taken are all age groups. All participants who participated in the study were given informed consent and have been briefed on the research procedure and possible unpleasant effects that may arise even if small. Sampling is conducted consecutively against all affordable populations that meet the study criteria. Materials and Methods Ninety-eight blood samples from malaria suspect patients at Tello Island Community Health Center, South Nias District of North Sumatra Province. The research was conducted with three methods, namely RDT SD Bioline Malaria-Ag / Pan Method, Microscope Method, Nested Polymerase Chain Reaction Method, two of which are Nested 1 Polymerase Chain Reaktion and Nested 2 Polymerase Chain Reaktion. The number of samples in this study 98 people. The number of women 50 patients (51%) more than in men 48 people (49%). The most ages were aged 15 years to 59 years, 41 patients (41.8%) and 5 years to 14 years old were 36 patients (36.7) and at least age above 60 years ie 6 patients (6.1 %). The most symptom is a fever that is 73 people (74,5%) and headache that are 71 patient (72,4%). Splenomegaly was found in 30 patients (30.6%). Population taken were a population with inclusion criteria based on clinical symptoms such as fever, headache, muscle aches, nausea, diarrhea, abdominal pain, appetite, anemia, and enlarged spleen, positive with RDT, Macroscopic P. falciparum, and P. vivax, P Ovale, P. malaria. From the findings with Nested PCR found a new type of P. Knowles. Results of examination with RDT, Microscopic, and Nested PCR in Plasmodium falciparum and Plasmodium vivax parasites. The highest sensitivity of examination (100%) on parasite examination was between RDT Pv and Nested PCR Pv. The highest specificity (100%) on parasitic examination was between RDT Pf with Nested PCR Pf and Microscopic Pf with Nested PCR Pf. The greatest predictive value of a positive test (+) (95.83%) was in RDT Pf with Nested PCR Pf and between Microscopic Pf and Nested PCR Pf. The largest predictive value of a negative test (-) (100%) is RDT Pv with Nested PCR Pv. The most infinitely predictable value of a positive test is between RDT Pf and Microscopic Pf. While the Predictive value of a test e most (0.31%) ie between RDT Pf with Nested PCR Pf and Microscopic Pf with Nested PCR Pf. Based on the results of research conducted found Plasmodium knowledge with new species in the Nias Islands of North Sumatra. These findings were obtained after confirmatory tests with Nest 2 PCR with a very small prevalence of 0.97%. This finding is an improvement in the search for new species of malaria causes especially in Sumatra and generally in Indonesia. Although the incidence is small, it can be detected because it uses the primary marker twice or Nest 2 PCR. This is possible because the primer can track new species down into genes or genotypes (properties that do not appear morphologically). Conclusions: Nested PCR can be used as a diagnostic for malaria falciparum, vivax, knowledge, mix infection and has been found Plasmodium Knowlesi in Nias Selatan District of North Sumatra Province although the prevalence is still low.

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