phylogenetic relationships amongst hev strains
DESCRIPTION
Phylogenetic relationships amongst HEV strains. Participants : Agnes Zotter Lambert Motilal Maria Montalvo Marissa Moses Robin Antoine. Hepatitis E Virus. Hepeviridae, Hepevirus genus Unenveloped RNA virus, 27-34nm in diameter +ve stranded RNA genome, 7.2 kb in size - PowerPoint PPT PresentationTRANSCRIPT
Phylogenetic relationships amongst HEV strains
Participants:•Agnes Zotter•Lambert Motilal•Maria Montalvo•Marissa Moses•Robin Antoine
Hepatitis E Virus• Hepeviridae, Hepevirus genus• Unenveloped RNA virus, 27-34nm in diameter• +ve stranded RNA genome, 7.2 kb in size• Very labile and sensitive
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Most outbreaks associated with faeceally contaminated drinking water.
Large epidemics have occurred in the Indian subcontinent, USSR, China, Africa and Mexico.
In the United States and other non-endemic areas, where there are no documented outbreaks of hepatitis E, a low prevalence of anti-HEV (<2%) has been found in healthy populations. The source of infection for these persons is unknown.
Minimal person-to-person transmission.
Hepatitis E - Epidemiologic
Features
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Incubation period: Average 40 days
Range 15-60 days Case-fatality rate: Overall, 1%-3%
Pregnant women, 15%-25%
Illness severity: Increased with age
Chronic sequelae: None identified
Hepatitis E - Clinical Features
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
HEV isolates have been grouped into four genotypes (1 to 4)
•Genotype 1 groups isolates found mainly in Asia and Africa
•Genotype 2 contains an isolate from an outbreak in Mexico and Africa
•Genotype 3 groups isolates found in the US & Europe
•Genotype 4 groups isolates found in China, Taiwan and Japan
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Hepatitis E Virus Genome
Research question
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
To determine the genotype identity of isolates found in Cuba
Possible ways of managing the disease
Methods:
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Sequences from RNA polymerase in GenBanK (NCBI)
13 hits
CUB9-1999 isolate chosen (241 bp linear RNA)Blastn: 30 entries chosen/E-value
Length +/- 10 nucleotides alignment
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Blastp Protein ID from initial selected isolate
ClustalX2 multiple alignmentAnalysis in Phylogenetic software
>gb|ACD39863.1| RNA-dependent RNA polymerase [Hepatitis E virus] Length=116
Score = 133 bits (334), Expect = 6e-37, Method: Compositional matrix adjust. Identities = 61/79 (77%), Positives = 69/79 (87%), Gaps = 0/79 (0%)
Query 1 CALFGPWFRAIEKAILALLPQGVFYGDAFDDTVFSATVAAAKASMVFENDFSEFDSTQNN 60 CALFGPWFRAIEK ILALLP +FYGDA++++VFSA ++ A +SMVFENDFSE ST NN Sbjct 9 CALFGPWFRAIEKE ILALLPPN IFYGDAYEES VFSAAISGAGSSMVFENDFSEDXSTLNN 68
Query 61 FSLGLECAIMEECGMPQWL 79 FSLGLEC IMEECGMPQWL Sbjct 69 FSLGLECVIMEECGMPQWL 87
Protein alignment (MSA):
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Results:
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
R
Nucleic Acids Research 2005 33(Web Server Issue):W553-W556; doi:10.1093/nar/gki494 . C-Y Lin*, F-K Lin, CH Lin, L-W Lai, H-J Hsu, S-H Chen and CA Hsiung
http://power.nhri.org.tw/
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Genotype 1
Genotype 3
Genotype 4
Genotype 2
DNA data
NJ analysis
Phylogenetic tree:
Protein Seq.data
NJ analysis
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Phylogenetic tree
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Phylogenetic tree: Phylip with DNA data JAP291
Cub25_08 Mex017
Fin969Fin973
Ind LonInd097
Mor494 Chn001
Chn363
Ind103
Chn457Hyder Cub19_99
Cub9_99NindCub2_05
Cub27_99Cub10_99
Fin971Fin967
US2SKor476
US1SKor466Fr757
Fr719Fr767
Fr787Fr785
Phylogenetic tree: Phylip/protein data
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
DNA Alignment without Cub25-2008
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Phylogenetic tree:
MEGA4/DNA data
Genotype 1
Genotype 2
Genotype 3
Genotype 4
Phylogenetic tree: MEGA/protein data
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Genotype 1
Genotype 2
Genotype 3
Genotype 4
HEV from human in Cuba were clustered in at least two genotypes.
Conclusion
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Genotype 1 with Asian and African isolates
Genotype 3 with American and European isolates from swine and human
A take-home message…
Avoid drinking water (and beverages with ice) of unknown purity, uncooked shellfish, and uncooked fruit/vegetables not peeled or prepared by traveler.
IG prepared from donors in Western countries does not prevent infection.
Unknown efficacy of IG prepared from donors in endemic areas.
Vaccine?
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Thank you for your attention!
UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.
Special thanks for the great teachers, who were very patient and helpful, who helped us to make this presentation better…
Dr. Urmila Kulkarni-Kale Dr. Jessica Kissinger Dr. Dinesh Gupta Dr. Arnab Pain
Dr. Vrijesh Tripathi