Groups 7: 1.Lailiya Vina Rochmatika (115090100111005) 2.Dita Fitriana Kusuma D. (115090101111003) 3.Dian Cahyaningtyas P. (115090107111007) MALARIA, ROLE.

Groups 7:

1. Lailiya Vina Rochmatika (115090100111005)

2. Dita Fitriana Kusuma D. (115090101111003)

3. 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 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

Malaria

parasite

Plasmodium

falciparum

P. vivax

P. malaria

e

P. ovale

4 TYPES OF MALARIA PARASITE

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 vivaxCause 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.

Plasmodium malariaeCause 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 ovaleThis type is generally a lot in Africa and the Western Pacific, causing ovale malaria.

Symptoms of

Malaria

Natural InfectionThis 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

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. AccidentalTransmission occurs through blood transfusion or through a syringe . Transmission through needle prevalent in drug addicts who use needles that are not sterile.

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

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

Serology is the scientific study of blood serum. In practice, the term usually refers to the diagnostic identification of antibodies in the serum.

Serology

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

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)

A Typical Indirect-Detection ELISA

A Typical Sandwich ELISA

ELISA have three detection for malaria:

1. Antibody detection (Seroepidemiology)

2. Antigen detection (Diagnosis)

3. Antigen detection (epidemiology)

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

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

Ratio Result Analysis

IgM < 0,9 Negative Nothing infection of malaria

IgM 0,9 – 1,1 ekuivocal Repeat test

IgM > 1,1 Positive Supposition new infection of malaria

IgG < 1,8 Negative Nothing secondary infection

IgG 1,8 – 2,2 Ekuivocal Repeat test

IgG > 2,2 positive Supposition new infection of malaria

Diagnosis of Humoral Respons with ELISA

PCR (Polymerase Chain Reaction) is the process of enzymatic synthesis to

amplify nucleotides in vitro

PCR (Polymerase Chain Reaction)

The primary basic cycle of PCR happens 30-35 cycle, including:1. Denaturation (950C), for 30 seconds. In this

step, the double helix of DNA separates into two units of single DNA.

2. 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.

3. 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

PCR can be used to identify:1. Genetic disorders2. Infection by viruses3. Early diagnose disorders such as

AIDS4. The genetic profile of forensic5. The application of biodiversity6. Biologycal evolution7. Direct genetic mutation8. Measure the quantification of mRNA

expression in cells or tissue

PCR (Polymerase Chain Reaction)

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

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 paper

More reliable for determining species in a mixed infection.

Can identify mutations- try to correlate to drug resistance

May have use in epidemiologic studies

Advantages

Parasitemia malaria not quantifiable Requires specialized equipment,

reagents, and training.

Disadvantages

Analysis of a PCR diagnostic test for species specific detection of Plasmodium DNA.

PCR was performed using nested primers.

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