8/8/2019 Rapid test for malaria

1/20

8/8/2019 Rapid test for malaria

2/20

INTRODUCTION

MALARIA INFECTION

life-threatening disease caused by parasites ofthespeciesPlasmodium

infectious

preventableand curable prevention: prophylactic antibiotic, doxycycline

treatment:combination of drugs knownasartemisinin-based

combinationtherapies (ACTs)

MOT:Bite ofinfected vectormosquito, female Anophelesminimus flavirotris

8/8/2019 Rapid test for malaria

3/20

SpeciesofPlasmodiumparasites: Can infect humans:

P. falciparum P.vivax

P.ovale P.malariae

INTRODUCTION

P. falciparum isthemost deadly.

P. falciparum and P. vivax arethemost prevalent.

Female Anophelesminimus flavirostris

8/8/2019 Rapid test for malaria

4/20

INTRODUCTION

Symptoms:first commonsymptoms whichusually appear10 to15 daysaftera personisinfected.

fever headache

chills vomiting

8/8/2019 Rapid test for malaria

5/20

INTRODUCTION

Ofutmostimportanceforbetterpatientmanagementand speedy recovery are:

Early detection

- Cerebralmalarialand drugresistanceassociated withfalciparum

malaria- Morbidity

Early differentiation

- course oftreatmentis dependent onthespecies

8/8/2019 Rapid test for malaria

6/20

DIAGNOSTIC TESTS

Blood Smear Gold standard

Rapid TestforMalaria Pan + Pv + Pf

FirstSign ParaView-3

Isarapid self-performing,qualitative, two sitesandwichimmunoassayutilizingwhole blood

8/8/2019 Rapid test for malaria

7/20

DIAGNOSTIC TESTS

Rapid Test for Malaria Pan + Pv + PfFirstSign ParaView-3

PARASITE DETECTION OF

P. falciparum P. falciparum specific histidine richprotein-2 (Pf. HRP-2)

P. vivax P. vivax specific parasitelactate dehydrogenase (pLDH)

P. ovale and P. malaria Panmalariaspecific parasite

lactate dehydrogenase(pLDH)

Detectionsystemis based on:

8/8/2019 Rapid test for malaria

8/20

isa watersoluble protein expressed onthered cellmembranesurface produced by theasexualstagesand gametocytes ofP.

falciparum

shownto remaininthe blood foratleast28 daysaftertheinitiation ofantimalarial therapy

solubleglycolytic enzyme produced by theasexualand sexualstages ofthelive

parasites itis presentinand released fromthe parasiteinfected

erythrocytes. Ithas beenfound inall 4 humanmalariaspecies, and differentisomers of pLDH foreach ofthe 4speciesexist

8/8/2019 Rapid test for malaria

9/20

PRINCIPLE

Immunochromatographic assay

-colloidalgold conjugate ofanti-HRP-2antibody,antiP. Vivax specific pLDH antibody and anti PanspecificpLDH antibody complexes HRP-2/corresponding pLDH

inthelysed sample.

-unreacted conjugatesalong withtherabbitantiseracolloidalgold conjugateand unbound complex(ifany),movesfurtheronthemebrane and are

subsequently immobilized by anti-rabbitantibodiescoated onthemembraneatthemembraneatthecontrolregion

8/8/2019 Rapid test for malaria

10/20

PRINCIPLE

8/8/2019 Rapid test for malaria

11/20

ANTIBODY AND ANTIGEN

ANTIBODY ANTIGEN

Monoclonalanti Pf.- HRP2antibody

monoclonalantiP.vivaxspecific pLDH antibody

monoclonalanti panspecific pLDH antibody

anti-rabbitantibody(control)

Pf. HRP2

P.vivax

Pan

8/8/2019 Rapid test for malaria

12/20

REAGENTS AND

MATERIALS USED1. TEST DEVICE:

Containsmembrane assembly

pre-dispensed with:

monoclonalanti- Pf. HRP2antibody-colloidalgold conjugate

monoclonalanti P.vivax specific pLDHantibody-colloidalgold conjugate

monoclonalanti panspecific pLDHantibody-colloidalgold conjugate

Rabbitglobulin colloidalgold conjugate

2. DESSICANT POUCH3. 5l SAMPLE LOOP

4. CLEARING BUFFER IN A DROPPER BOTTLE

5. PACKAGE INSERT

8/8/2019 Rapid test for malaria

13/20

DESCRIPTIONS OF THE TEST

Canspecifically detectand differentiate Pf & Pv-which constituteabout 90% ofthetotalmalaria occurrence

globally.

Good clinicalsensitivity and specificity >98% forP.falciparum malariae-dueto HRP-2 based detection.

Ableto monitorsuccess ofanti-malarialtherapy-through Pan band.

8/8/2019 Rapid test for malaria

14/20

LIMITATIONS OF THE TEST

Cannotspecifically detectand differentiateP.malariae andP.ovale.

HRP-2 isnotsecreted ingametogony stage. Hence,in carriers, the HRP-2may beabsent.

8/8/2019 Rapid test for malaria

15/20

8/8/2019 Rapid test for malaria

16/20

PROCEDURES

8/8/2019 Rapid test for malaria

17/20

8/8/2019 Rapid test for malaria

18/20

INTERPRETATION OF RESULTS

8/8/2019 Rapid test for malaria

19/20

INTERPRETATION OF RESULTS

Negative (-)A negativeresultindicatestheabsence ofany malarialparasiteinthesample

Positive (+):

A positiveresult onthe Pfregionindicatesthe presence ofP. falciparum infection

A positiveresult onthe Pvregionindicatesthe presence ofP.vivax infection

A positiveresult onthe Panregion butanegativeresult oneitherPfand Pvregionsindicatetheinfection of othermalarial parasitespecies ( P.malariae and P.ovale).

A positiveresult onalltheregionsindicateamixed malarial

parasiteinfection

8/8/2019 Rapid test for malaria

20/20

MANNER OF REPORTING

Negative (-)Only one pink/pink-purple band appearsatthe controlregionC

Positive (+):

1. P. falciparum malariaInadditionto the control band, a pink/purple band appearsat

the Pfand Panregionsrespectively

2. P. vivax malariaInadditionto the control band, a pink/purple band appearsat

Pvand Panregionsrespectively

3. OtherspeciesInadditionto the control band, one pink/purple band appears

only atthe Panregion

4. Mixed infectionInadditionto the control band, a pink/purple band appearsat

Pf, Pvand Panregions