INDIA /BRAZIL / SOUTH AFRICA TRI-LATERAL COLLABORATIVE PROGRAM ON SCIENCE AND TECHNOLOGY II Forum IBSA on Malaria, Teresina – PI, BR, March, 7th - 8th, 2006 Hernando A del Portillo, PhD Departamento de Parasitologia Instituto de Ciências Biomédicas Universidade de São Paulo http://clinmaldb.usp.br/hadelportillo

Diagnosis, Cell and Molecular Biology

DIAGNOSIS  

Giemsa-staining Malaria Diagnosis Brazil 1960-2005

External Evaluation of Giemsa-staining diagnosis in reference laboratories from the State of Pará Dra. Marinete Póvoa State of Pará more than 100.000 cases/2005 11 participating laboratories Five different reference slides: P. falciparum, P. vivax, mixed infection, P. vivax + T. cruzi, negative control 73% of the labs diagnosed correctly all slides, remaining 27% miss-diagnosed 1 slide. None of the labs identified T. cruzi. 5 labs have adequate training; 4 labs require re-training and 2 labs require capacitance.

Multi-Center Assesstment of Immuno-chromatographic Methods Coordinated: Prof. Cor Jésus Fernandes Fontes

Giemsa Staining Total No of Patients : 539 Resuts given by Manaus, Porto Velho, Macapá e Cuiabá. Giemsa Staining F V N Total 87 2 89 OPTIMAL V* 104 4 9 333 346 T otal 91 113 335 539 * = Positive results of other humana malaria parasite species, here in considered positive for P. vivax Sensitivity P.falciparum = 95,6% P.vivax = 92,0 Specificity = 99,5% = 100,0% Overall sensitivity for malaria diagnosis = 93,6% Overall specificity for malaria diagnosis = 99,4%

PCR

NESTED-PCR

NESTED-PCR Evaluation of negativation and detection of three human malaria parasite species: Marinete Póvoa Blood bank surveys Ricardo Machado MF Ferreira-Da-Cruz Mixed Infections Graça Alecrim Asymptomatics LH Pereira da Silva

Nested-PCR and Malaria Diagnosis at FMT/AM Nested-PCR detected 14.5% of mixed infections as opossed to 1,8% detected by Giemsa staining

Asymptomatic patients

ELISA Determination of the cutoff value for IgG-ELISA-P. vivax MSP1-19, using the Receiver Operating Characteristic (ROC) Curve. Sandra do Lago Moraes-Avila & Antonio Walter Ferreira

Nogueira P, Alves F et al

Giemsa-staining Malaria Diagnosis Brazil 1960-2005

MOLECULAR AND CELL BIOLOGY  

Polymorphims of Molecular Markers - Parasites P. vivax Duffy Binding Protein (de Brito, Moreira, Carvalho) P. falciparum reticulocyte binding-like homologues (PfRh1-4) (Mariano Zalis) P126 protein (Ferreira-da-Cruz, Banic, Carvalho, Ribeiro) Glycophorin B (GBP), HFE (Ricardo Machado) MSP1, MSP2 (Marcelo Urbano)

Baum et al PLoS Path 2005

Zalis, M (unpublished)

Microssatelites Plasmodium vivax Cor Jesús Fernandes Fontes, Alvaro Couto, Luzia H Carvalho Cristiana FA Brito 2006

Polymorphisms - Human Molecular Markers and Associations with Malaria Infection CR-1, MBL, TLR, NOS2, TNF-a, IFN-b, IL-4, IL-10 CD36, ICAM-1, Cytochrome P450, GST (Sandra do Lago Moraes)

CSP MSP119

Mosquitoes and cDNA libraries cDNA library of A. darlingi W. P. Tadei