Abstract
Leishmaniasis, manifesting on it`s different clinical forms is endemic in different regions of the Sudan. diagnosis of the disease in the Sudan is usually done using simple methods such as microscopical examination of slit smirs, Histological sections and cultures. Serological diagnosis using enzymes linked immunosorbent assay (ELISA)- and direct agglutination test (DAT) are sometimes used as more sensitive tool has thrown light on the epidemiology of the disease in the Sudan. This study was conducted on 126 subjects to identify the parasites-causing the different clinical manifestations, to determine the genetic diversity of different isolates of Lieshmania- and to detect parasites in the peripheral blood of subjects from highly endemic foci. The study population consisted of 7 with suspected VL, 12 with suspected ML, 14 with suspected PKDL, 2 with sporotrichoid CL and 89 healthy games wardens and army soldiers from highly endemic foci. Parasites were cultured in biphasic medium and subcultured in liquid medium until mass production was stabilized. Extraction of DNA was done using three methods which were phenol/ chloroform/ isoamylalcohol, K buffer and proteinase K as well as lysing of the parasite with distilled water. The KDNA was amplified using species namely AJSI and DeB8. The products were
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Hashim, Amna Osman Yousif
[1]
- Department of Zoology, Faculty of Science, University of Khartoum, Khartoum (Sudan)
Citation Formats
Hashim, Amna Osman Yousif.
The Polymerase chain reaction as a tool of molecular diagnosis of Leishmania infection in the Sudan.
Sudan: N. p.,
1997.
Web.
Hashim, Amna Osman Yousif.
The Polymerase chain reaction as a tool of molecular diagnosis of Leishmania infection in the Sudan.
Sudan.
Hashim, Amna Osman Yousif.
1997.
"The Polymerase chain reaction as a tool of molecular diagnosis of Leishmania infection in the Sudan."
Sudan.
@misc{etde_320124,
title = {The Polymerase chain reaction as a tool of molecular diagnosis of Leishmania infection in the Sudan}
author = {Hashim, Amna Osman Yousif}
abstractNote = {Leishmaniasis, manifesting on it`s different clinical forms is endemic in different regions of the Sudan. diagnosis of the disease in the Sudan is usually done using simple methods such as microscopical examination of slit smirs, Histological sections and cultures. Serological diagnosis using enzymes linked immunosorbent assay (ELISA)- and direct agglutination test (DAT) are sometimes used as more sensitive tool has thrown light on the epidemiology of the disease in the Sudan. This study was conducted on 126 subjects to identify the parasites-causing the different clinical manifestations, to determine the genetic diversity of different isolates of Lieshmania- and to detect parasites in the peripheral blood of subjects from highly endemic foci. The study population consisted of 7 with suspected VL, 12 with suspected ML, 14 with suspected PKDL, 2 with sporotrichoid CL and 89 healthy games wardens and army soldiers from highly endemic foci. Parasites were cultured in biphasic medium and subcultured in liquid medium until mass production was stabilized. Extraction of DNA was done using three methods which were phenol/ chloroform/ isoamylalcohol, K buffer and proteinase K as well as lysing of the parasite with distilled water. The KDNA was amplified using species namely AJSI and DeB8. The products were analysed on 1.5% agarose gel-and were visualized and photographed with U.V. transilluminator and camera. Characteristic bands of 700 and 800 b.p corresponding to the full length of mini circle of L.major and L.donovani respectively were obtained on amplification of KDNA from patients with VL and CL. In some cases lower bands of 400 and 500 b.p PKDL and multiple bands for sporotrichoid CL.Leishmania DNA was detected from the conjucativa of the eye of a patient with PKDL. The genetic diversity of Leishmania parasite was determined by digesting PCR products from PKDL, sporotrichoids CL and VL patients. Different patterns were produced for each digesting product. This result indicates that there is heterogeneity among isolates producing products of 800 bp. The data from the Dinder national park showed that Leishamanin skin testing and K39 dipstick may be useful as endemicity marker in these areas. PCR detected L.donovani on patients peripheral blood who displayed a wide clinical spectrum including healthy individual of both Leishmania negative and positive, active kala-zar cases well as previous kala-zar patients. The study has shown that different isolates of Leishmania cause clinical forms and there is an indication of heterogeneity among isolates causing VL, PKDA and sporotrichoid CL.(Author) 119 ref. , 2 tabs. , 12 figs.}
place = {Sudan}
year = {1997}
month = {Jun}
}
title = {The Polymerase chain reaction as a tool of molecular diagnosis of Leishmania infection in the Sudan}
author = {Hashim, Amna Osman Yousif}
abstractNote = {Leishmaniasis, manifesting on it`s different clinical forms is endemic in different regions of the Sudan. diagnosis of the disease in the Sudan is usually done using simple methods such as microscopical examination of slit smirs, Histological sections and cultures. Serological diagnosis using enzymes linked immunosorbent assay (ELISA)- and direct agglutination test (DAT) are sometimes used as more sensitive tool has thrown light on the epidemiology of the disease in the Sudan. This study was conducted on 126 subjects to identify the parasites-causing the different clinical manifestations, to determine the genetic diversity of different isolates of Lieshmania- and to detect parasites in the peripheral blood of subjects from highly endemic foci. The study population consisted of 7 with suspected VL, 12 with suspected ML, 14 with suspected PKDL, 2 with sporotrichoid CL and 89 healthy games wardens and army soldiers from highly endemic foci. Parasites were cultured in biphasic medium and subcultured in liquid medium until mass production was stabilized. Extraction of DNA was done using three methods which were phenol/ chloroform/ isoamylalcohol, K buffer and proteinase K as well as lysing of the parasite with distilled water. The KDNA was amplified using species namely AJSI and DeB8. The products were analysed on 1.5% agarose gel-and were visualized and photographed with U.V. transilluminator and camera. Characteristic bands of 700 and 800 b.p corresponding to the full length of mini circle of L.major and L.donovani respectively were obtained on amplification of KDNA from patients with VL and CL. In some cases lower bands of 400 and 500 b.p PKDL and multiple bands for sporotrichoid CL.Leishmania DNA was detected from the conjucativa of the eye of a patient with PKDL. The genetic diversity of Leishmania parasite was determined by digesting PCR products from PKDL, sporotrichoids CL and VL patients. Different patterns were produced for each digesting product. This result indicates that there is heterogeneity among isolates producing products of 800 bp. The data from the Dinder national park showed that Leishamanin skin testing and K39 dipstick may be useful as endemicity marker in these areas. PCR detected L.donovani on patients peripheral blood who displayed a wide clinical spectrum including healthy individual of both Leishmania negative and positive, active kala-zar cases well as previous kala-zar patients. The study has shown that different isolates of Leishmania cause clinical forms and there is an indication of heterogeneity among isolates causing VL, PKDA and sporotrichoid CL.(Author) 119 ref. , 2 tabs. , 12 figs.}
place = {Sudan}
year = {1997}
month = {Jun}
}