Dita Fitriana Kusuma D 115090101111003 Dian Cahyaningtyas P 115090107111007 MALARIA ROLE OF SEROLOGY ELISA AND PCR Malaria is a disease caused by parasites of the genus Plasmodium including protozoa through the mediation group puncture bite mosquito ID: 1043633
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1. Groups 7:Lailiya Vina Rochmatika (115090100111005)Dita Fitriana Kusuma D. (115090101111003)Dian Cahyaningtyas P. (115090107111007)MALARIA, ROLE OF SEROLOGY: ELISA AND PCR
2. Malaria is a disease caused by parasites of the genus Plasmodium including protozoa through the mediation group puncture (bite) mosquito Anopheles spp.Malaria is one of the world's problems are the main public health. Malaria spreads in many countries, especially in Asia, Africa, and America Latin.MALARIA
3. 4 TYPES OF MALARIA PARASITE
4. Plasmodium falciparum Falciparum malaria malignant (malaria tropica) causes fever patients every day. The incubation period of malaria or the time between the mosquito bite and the appearance of clinical symptoms about 7-14 days.Plasmodium vivax Cause disease vivax malaria (benign tertian malaria). The incubation period of malaria or the time between the mosquito bite and the appearance of clinical symptoms about 8-14 days.
5. Plasmodium malariae Cause disease malaria quartana. The incubation period of malaria or the time between the mosquito bite and the appearance of clinical symptoms about 7-30 days.Plasmodium ovale This type is generally a lot in Africa and the Western Pacific, causing ovale malaria.
6. Symptoms of Malaria
7. Natural Infection This transmission occurs through the bite of an infective female Anopheles mosquito . Mosquitoes bite people sick with malaria parasite will then participate sucked the blood of patients with malaria . Parasite in the mosquito body will grow and multiply , then a mosquito bites a healthy person, the parasite transmitted through the bite to other people.Transmission of Malaria
8. Unnatural Infectiona. Congenital Malaria Occurs in newborns because her mother suffered from malaria . Due to abnormalities in the placental barrier so that there is no barrier of infection from mother to baby.b. Accidental Transmission occurs through blood transfusion or through a syringe . Transmission through needle prevalent in drug addicts who use needles that are not sterile.
9. Benign malaria can be given orally. Whereas malignant malaria have clinical symptoms of bleeding should be observed in the hospital with intravenous medication.Treatment of Malaria
10. 1. Microscopic Diagnosis a. Blood smear b. Fluorescent Microscopy c. Quantitive Buffycoat (QBC)2. Antigen Detection a. Immunochromatographic Dipstick : RDT3. Serology a. IFA b. ELISA4. Molecular Diagnosis a. PCR Laboratory Diagnosis of Malaria
11. Serology is the scientific study of blood serum. In practice, the term usually refers to the diagnostic identification of antibodies in the serum.Serology
12. Serologycal test is a test using serum to diagnose some disorders such as malaria. Using this methods it will be possible to do research to the reaction of antigen-antibody (Ag-Ab) in vitro. This method is based on the process of precipitation, agglutination, or complemen activation.Serologycal Test
13. is a widely used biochemical technique for the detection of an antigen in a sample.ELISA assays are frequently used in viral diagnostics , for example in detecting cases of HIV infection. And than used diagnostic of malaria. ELISA (Enzyme-linked Immunosorbent Assay)
14. A Typical Indirect-Detection ELISA
15. A Typical Sandwich ELISA
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18. ELISA have three detection for malaria:1. Antibody detection (Seroepidemiology)2. Antigen detection (Diagnosis)3. Antigen detection (epidemiology)
19. ELISA used to identify the epidemiology of malaria-infected mosquitoes. The use of monoclonal antibodies specific for protein of Circum Sporozoite Plasmodium knowlesi. This monoclonal antibody used the solid phase and conjugated with an enzyme, as signifying the presence of protein in the sporozoite Circum mosquito homogenates that incubated in microplate wells.ELISA for Diagnosis of Malaria
20. Circum sporozoite protein (CSP) is an important antigen found on the surface of sporozoites, causing a role in antibody-mediated protection against parasites. Monoclonal antibodies produced to the antigen specificity was determined. By antigen capture ELISA is a useful method for rapidly detecting specific protein antigen as well as in homogenates of mosquitoes (mosquitoes scours pul).ELISA for Diagnosis of Malaria
21. RatioResultAnalysisIgM < 0,9NegativeNothing infection of malariaIgM 0,9 – 1,1ekuivocalRepeat testIgM > 1,1PositiveSupposition new infection of malariaIgG < 1,8NegativeNothing secondary infectionIgG 1,8 – 2,2EkuivocalRepeat testIgG > 2,2positiveSupposition new infection of malariaDiagnosis of Humoral Respons with ELISA
22. PCR (Polymerase Chain Reaction) is the process of enzymatic synthesis to amplify nucleotides in vitroPCR (Polymerase Chain Reaction)
23. The primary basic cycle of PCR happens 30-35 cycle, including:Denaturation (950C), for 30 seconds. In this step, the double helix of DNA separates into two units of single DNA.Annealing (550C-600C), for 30 seconds. In this step happens to describe the binding of single DNA probes, the temperature of this step is defined by the primary.Extension (720C), in this step, polymerase happens to form a new single DNA. The duration depends both on the DNA polymerase used and on the length of the DNA fragment to be amplified.The Primary Basic Cycle of PCR
24. PCR can be used to identify:Genetic disordersInfection by virusesEarly diagnose disorders such as AIDSThe genetic profile of forensicThe application of biodiversityBiologycal evolutionDirect genetic mutationMeasure the quantification of mRNA expression in cells or tissuePCR (Polymerase Chain Reaction)
25. Molecular teqnique to identify parasite genetic material of malaria in Plasmodium DNA.Uses whole blood collected in anticoagulated tube (200 μl) or directly onto filter paper (5 μl)PCR for Diagnosis Malaria
26. PCR is a reference method. It is at least 10- fold more sensitive than microscopy. - Threshold detection * 0,1 parasite of malaria / μl if whole blood in tube * 2 parasite of malaria / μl if using filter paperMore reliable for determining species in a mixed infection.Can identify mutations- try to correlate to drug resistanceMay have use in epidemiologic studiesAdvantages
27. Parasitemia malaria not quantifiableRequires specialized equipment, reagents, and training.Disadvantages
28. Analysis of a PCR diagnostic test for species specific detection of Plasmodium DNA. PCR was performed using nested primers.
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