378 17th International Congress on Infectious Diseases / International Journal of Infectious Diseases 45S (2016) 1477
Type: Poster Presentation
Final Abstract Number: 43.071
Session: Poster Session III
Date: Saturday, March 5, 2016
Time: 12:45-14:15
Room: Hall 3 (Posters & Exhibition)
HLA determinants of susceptibility and
protection to L. donovani: in silico analysis
N. Samaranayake , D. Fernando, N. Karunaweera,
V. Dissanayake
Faculty of Medicine, University of Colombo,
Colombo, Sri Lanka
Background: Sri Lanka is endemic for leishmaniasis caused by
Leishmania donovani Mon 37. Localised cutaneous leishmaniasis
(LCL) is the predominant presentation with a few reports of visceral
disease. Limited differences in the two strains causing cutaneous
and visceral phenotypes (1) suggest concomitant operation of host
factors. A case-control association study conducted by us suggested
HLA*A68 and HLA*B07 to confer protection to LCL (submitted for
publication). The objective of this study was to further assess these
ndings by predicting MHC-I binding for nonameric peptides in
identied L. donovani sequences.
Methods & Materials: Protein coding genes of L. dono-
vani reference strain BPK282A1 were retrieved from TryTripDB.
Protein sequences of genes (LdBPK 365490.1, LdBPK 303700.1,
LdBPK 261610.1 and LdBPK 220670.1) shown to be over expressed
in L. donovani strains causing cutaneous and visceral disease in the
country were identied. The MHC-I binding predictions for these
peptides were made using the IEDB analysis resource Consensus
tool. Binding of HLA alleles associated with LCL were compared with
the 5 most common alleles at these loci (threshold percentile rank
<=1). Common alleles were selected according to data available for
the local population.
Results: The total number of epitopes predicted for selected
alleles at HLA-A and HLA-B loci were 70 and 52 respectively. At
the HLA-A locus, A*68 had the highest number of predicted epi-
topes (n = 19). Majority of these (n = 13) were on adaptin protein
coded by LdBPK 365490.1 and 11 of them were high afnity binders
(IC50 <50 nM). At the HLA-B locus, B*07 was predicted to bind
the most number of epitopes (n = 14) with a similar distribution
in proteins encoded by LdBPK 303700.1 and LdBPK 365490.1. One
epitope each on A2 proetein was predicted to bind HLA-A*02 and
HLA-B*57. No epitopes on this protein were predicted for HLA*A68
and HLA*B07.
Conclusion: Alleles which had an apparent protective effect
according to our previous ndings consistently demonstrated abil-
ity to bind more epiotpes, of proteins encoded by genes over
expressed in parasite strains causing LCL. The immune responses in
this host-parasite combination should be studied in a larger sample,
in a HLA restricted setting, to elucidate any protective mechanisms.
http://dx.doi.org/10.1016/j.ijid.2016.02.810
Type: Poster Presentation
Final Abstract Number: 43.072
Session: Poster Session III
Date: Saturday, March 5, 2016
Time: 12:45-14:15
Room: Hall 3 (Posters & Exhibition)
Evaluation of loop mediated isothermal
amplication for the diagnosis of amoebic liver
abscess
R. Sehgal 1, , S. Khunger 2 , A. Duseja 2 , D. Handa 3 ,
R. Singh 3
1 Post Graduate Institute of Medical Education and
Research, Chandigarh, Chandigarh, India
2 PGIMER, Chandigarh, India
3 GMCH-32, Chandigarh, India
Background: Entamoeba histolytica is an important cause of
amoebic liver abscess (ALA) in humans, particularly in developing
countries. Molecular techniques are increasingly used for the detec-
tion of infectious agents in clinical samples with high sensitivity and
specicity. The present study has been planned to investigate the
utility of Loop mediated isothermal amplication for the detection
of E. histolytica/E. dispar in suspected cases of amoebic liver abscess.
Methods & Materials: A total of 138 pus samples were collected
from the suspected patients of ALA who attended emergency med-
ical OPD/Liver clinic of Postgraduate Institute of Medical Education
and Research, Chandigarh/Government Medical College and Hospi-
tal, Sector-32 Chandigarh from November 2013 to November 2014.
Pus samples of liver abscess were obtained by ultrasound guided
aspiration and subjected for diagnostic evaluation using Loop medi-
ated isothermal amplication and Real time PCR for the detection
of E. histolytica/E. dispar. ELISA was performed for the detection of
anti-Entamoeba IgG antibodies on the corresponding serum sam-
ples.
Results: The M:F ratio of the recruited patients was 16.1:1.
Loop mediated isothermal amplication detected E. histolytica DNA
among 58 of 133 (43%) pus samples as compared to Real time PCR
which detected 28 of 133 (21.66%) pus samples. Anti-Entamoeba
antibodies were detected by ELISA in serum of 99% of cases.
Conclusion: Thus, loop mediated isothermal amplication
proves to be a promising diagnostic tool in providing direct
detection and conrmatory evidence of amoebic liver abscess in
comparison to Real time PCR and antibody detection by ELISA.
http://dx.doi.org/10.1016/j.ijid.2016.02.811