Malaria Slide Reading

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1 Malaria Slide Reading La Lecture des lames pour le diagnostic du paludisme Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

2 Content Contenu Malaria in general Le paludisme en général Preparation of Slides La préparation de lames Coloration -> Giemsa Staining La coloration avec Giemsa Microscope handling Maintenance du microscope Counting methods Les méthodes pour compter Differentiation of Species La distinction des espèces Quiz Quiz Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

3 Malaria Paludisme Parasitic infection Infection parasitaire Transmitted by the female Anopheles mosquito Transmis par le moustique Anophèle femelle 4 species which infect humans: Plasmodium falciparum, P. malariae, P. vivax, P. ovale 4 espèces qui infectent l Homme: Plasmodium falciparum, P. malariae, P. vivax, P. ovale Major symptoms: high fever, chills, headache, body pain Les symptômes majeurs: fièvre, frissons, maux de tête, courbatures Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

4 Life cycle of plasmodium Le cycle de plasmodium Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

5 Preparation Préparation Use of venous blood (EDTA) or capillary blood (fingerprick) Utilisation du sang veineux (EDTA) ou capillaire Thin and thick smears Faites les lames rapidement Thin smear: differentiation of species Thick smears: quantification of parasites (Routine) Important : Identification! Goutte épaisse et frottis Frotti: Pour différentier des espèces Goutte épaisse: quantification des parasites (routine) Important: Identification! Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

6 Slides - Lames Thick Smear Goutte épaisse Thin Smear- frottis 2011 by American Society of Hematology Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

7 Giemsa Staining Giemsa Coloration Giemsa solution 20 % concentration: 20 ml of Giemsa 80 ml of buffer ph 7,2 filtration Solution should be not older than 4-5 hours Staining for 20 minutes Washing with clean water Drying in dry incubator (~40 C) or air-dry Solution de Giemsa: concentration à 20 %: 20 ml de Giemsa 80 ml de buffer ph 7,2 filtration La solution ne doits pas être vieille de plus de 4-5 heures Coloration pour 20 minutes Nettoyer avec de l eau claire Sécher dans l incubateur sec (~40 C) ou à l air Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

8 Microscope Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme 10x objective 100x objective + immersion oil Afterwards cleaning with Wundbenzin Look through the slide systematically L objective de 10x L objective de 100x avec l huile immersion Après nettoyer l objective avec du benzène Lire systématiquement la lame

9 Methods Lambaréné method Méthodes La méthode de Lambaréné Counting of microscope fields and parasites Compter les champs du microscope et les parasites negative: 100 fields négatif: 100 champs <5 parasites: 100 fields <5 parasites: 100 champs 5-50 parasites: 30 fields 5-50 parasites: 30 champs 50+ parasites: 20 fields 50+ parasites: 20 champs Calculation of parasitemia: number of parasites x microscope factor number offields Parasites/µl Calcul de la parasitémie: nombre de parasites nombre de champs parasites/µl facteur de microscope Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

10 Methods WHO Method Méthodes La méthode de OMS Counting of leukocytes and parasites Compter les leucocytes et des parasites negative: 500 Leukocytes négative: 500 Leucocytes 0-10 parasites: 500 Leukocytes 0-10 parasites: 500 Leucocytes 10+ parasites: 200 Leukocytes 10+ parasites: 200 Leucocytes Calculation of parasitemia: Calcul de la parasitémie: Number of Parasites x 8000 Number of Leukocytes Nombre de parasites x 8000 Nombre de Leucocytes Parasites/µl Parasites/µl Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

11 What do you see? Que voyez-vous? Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

12 Plasmodium Species/Espèces de plasmodium Plasmodium falciparum most frequent La plus courante Severe form Les cas grâves Plasmodium malariae Less frequent La moins courante Plasmodium ovale Rare species Espèce rare Frequent in Africa Courante en Afrique Plasmodium vivax Not frequent in Africa Pas courante en Afrique Biggest species (size) La plus grosse espèce (taille) Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

13 Plasmodium Species Differentation

14 Differentation des espèces de Plasmodium

15 Plasmodium Species Differentation

16 Differentation des espèce de Plasmodium

17 Quiz Which parasites or species do you see? Quel parasites ou espèce voyez vous? Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

18 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

19 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

20 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

21 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

22 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

23 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

24 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

25 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

26 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

27 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

28 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

29 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

30 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

31 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

32 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

33 Malcolouration LEUCOCYTES Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

34 Literature / Littérature BASIC MALARIA MICROSCOPY/ DIAGNOSTIC MICROSCOPIQUE DU PALUDISME Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

35 Malaria Slide Reading Training/La Lecture des lames pour le diagnostic du paludisme

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