RESUMO
Comparam-se neste trabalho resultados de identificaçäo de espécie de Plasmodium obtidos com a microscopia convencional de gotas espessas coradas pelo Giemsa, realizada no local, e um protocolo semi-aninhado de reaçäo em cadeia da polimerase (PCR) em 96 pacientes maláricos de Rondônia, Amazônia Ocidental Brasileira. Infecçöes envolvendo mais de uma espécie foram detectadas através de PCR em 30 por cento dos pacientes, mas nenhum caso havia sido encontrado pelo exame de gotas espessas. Além disso, encontraram-se infecçöes gor P. malariae por PCR em 9 dos 96 pacientes (10 por cento), mas nenhuma havia sido encontrada pelos microscopistas locais. Discute-se o potencial impacto de erros de identificaçäo de espécie sobre o tratamento e o controle da malária
Assuntos
Humanos , Corantes Azur , Plasmodium malariae/isolamento & purificação , Reação em Cadeia da Polimerase , Malária/diagnóstico , Malária/epidemiologia , Malária/prevenção & controleRESUMO
The merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum codes for a major asexual blood-stage antigen currently proposed as a major malaria vaccine candidate. The protein, however, shows extensive polymorphism, which may compromise its use in sub-unit vaccines. Here we compare the patterns of allelic diversity at the MSP-1 locus in wild isolates from three epidemiologically distinct malaria-endemic areas: the hypoendemic southwestern Brazilian Amazon (n=54), the mesoendemic southern Vietnam (n=238) and the holoendemic northern Tanzania (n=79). Fragments of the variable blocks 2, 4a, 4b and 6 or 10 of this single-copy gene were amplified by the polymerase chain reaction, and 24 MSP-1 gene types were defined as unique combinations of allelic types in each variable block. Ten different MSP-1 types were identified in Brazil, 23 in Vietnam and 13 in Tanzania. The proportion of genetically mixed infections (isolates with carrying more one MSP-1 version) ranged from 39 per cent in Brazil to 44 per cent in Vietnam and 60 per cent in Tanzania. The vast majority (90 per cent) of the typed parasite populations from Brazil and Tanzania belonged to the same seven most frequent MSP-1 gene types. In contrast, these seven types corresponded to only 61 per cent of the typed parasite populations from Vietnam. Non-random associations were found between allelic types in blocks 4a and 6 among Vietnamese isolates, the same pattern being observed in independent studies performed in 1994, 1995 and 1996. These results suggest that MSP-1 is under selective pressure in the local parasite population. Nevertheless, the finding that similar MSP-1 type frequencies were found in 1994 and 1996 argues against the prominence of short-term frequency-dependent immune selection of MSP-1 polymorphisms. Non-random associations between MSP-1 allelic types, however, were not detected among isolates from Brazil and Tanzania. A preliminary analysis of the distribution od MSP-1 gene types per host among isolates from Tanzania, but not among those from Brazil and Vietnam, shows significant deviation from that expected under the null hypothesis of independent distribution of parasites carrying different gene types in the human hosts. Some epidemiological consequences of these findings are discussed.