Analysis Of The Nagt Gene Diversity Among Leishmania Isolates From Sudan
Analysis Of The Nagt Gene Diversity Among Leishmania Isolates From Sudan
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Date
2007-09
Authors
Mossalami, Gihan
M. Mukhtarss, Maowia
M. Elamin, Dr. Elwaleed
Journal Title
Journal ISSN
Volume Title
Publisher
University of Khartoum
Abstract
Sixty one Leishmania isolates representing different clinical forms of leishmaniasis
(visceral leishmaniasis and HIV/Leishmania coinfection, cutanous leishmaniasis, post
kalazar dermal leishmaniasis and mucosal leishmaniasis) from Sudan were analysed
using the polymerase chain reaction and the analysis of the restriction fragment length
polymorphism (RFLP) of intra geneic region of (NAGT gene) and sequencing technique.
PCR amplification of (NAGT) gene of sixty one leishmania isolates resulted in the
amplification of a DNA fragment of 1.4kb.
The amplification of the (NAGT) gene coding region followed by restriction fragment
length polymorphism (PCR-RFLP) differentiated between Sudanese leishmania isolates
and clustered them as two groups; leishmania donovani complex and leishmania major,
but did not differentiate between the members of leishmania donovani complex. The
Acc1 restriction enzyme findings revealed the high conservation of this region.
The dendrogram of unweighted pair group method with arithmetic average (UPGMA)
and Neighbour-Joining (NJ) was constructed from the genetic distances derived from
PCR-RFLP data. The topology of the NJ and UPGMA trees were very similar.
DNA sequencing and sequence alignment analysis of (NAGT) gene differentiated the
leishmania donovani complex from leishmania major, but did not differentiate the
member's leishmania donovani complex, and agreed with the result obtained by
PCR⁄RFLP analysis. The (NJ) tree developed based on sequence alignment segregate
(NAGT) gene in to two groups having a one common ancestor.
In conclusion the two trees (from PCR-RFLP and from sequencing analysis) indicated
that there was no genetic heterogeneity among tested Sudanese isolates, and no
correlation between the PCR- RFLP polymorphic pattern, NAGT gene sequence and the
clinical presentation of the human leishmaniasis.
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Keywords
LEISHMANIASIS,Cutaneous leishmaniasis (CL),Mucocutaneous leishmaniasis (MCL),Visceral leishmaniasis (VL),Leishmania species,Leishmanin skin test (LST),Enzyme linked immunosorbent assay (ELISA),Indirect immunofluorecent antibody (IFAT)