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Phylogenetic relationships amongst HEV strains

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

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Phylogenetic relationships amongst HEV strains

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  1. Phylogenetic relationships amongst HEV strains • Participants: • AgnesZotter • LambertMotilal • Maria Montalvo • MarissaMoses • RobinAntoine

  2. 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.

  3. Hepatitis E - Epidemiologic Features • 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. UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  4. Hepatitis E - Clinical Features • 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 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  5. UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  6. Hepatitis E Virus Genome • HEVisolateshavebeengroupedintofourgenotypes (1 to 4) • Genotype 1 groupsisolatesfoundmainly in Asia and Africa • Genotype 2 containsanisolatefromanoutbreak in Mexico and Africa • Genotype 3 groupsisolatesfound in the US & Europe • Genotype 4 groupsisolatesfound in China, Taiwan and Japan UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  7. Research question To determine the genotype identity of isolates found in Cuba Possible ways of managing the disease UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  8. Methods: Sequencesfrom RNA polymerase in GenBanK (NCBI) 13 hits CUB9-1999 isolatechosen (241 bp linear RNA) Blastn: 30 entrieschosen/E-value Length +/- 10 nucleotidesalignment UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  9. BlastpProtein ID frominitialselectedisolate >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 ClustalX2 multiplealignment Analysis in Phylogeneticsoftware UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  10. Protein alignment (MSA): UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  11. Results: 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.

  12. Phylogenetic tree: Genotype 4 DNA data NJ analysis Genotype2 Genotype 3 Genotype 1 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  13. Protein Seq. data Phylogenetic tree NJ analysis UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  14. Phylogenetic tree: Phylip with DNA data JAP291 Cub25_08 Mex017 Fin969 Fin973 Ind LonInd097 Mor494 Chn001 Chn363 Ind103 Chn457 Hyder Cub19_99 Cub9_99 Nind Cub2_05 Cub27_99 Cub10_99 Fin971 Fin967 US2 SKor476 US1 SKor466 Fr757 Fr719 Fr767 Fr787 Fr785 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  15. Phylogenetic tree: Phylip/protein data UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  16. DNA Alignment without Cub25-2008 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  17. Phylogenetic tree: MEGA4/DNA data Genotype 1 Genotype 2 Genotype 4 Genotype 3 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  18. Phylogenetic tree: MEGA/protein data Genotype 1 Genotype 2 Genotype 4 Genotype 3 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  19. Conclusion HEV fromhuman in Cuba wereclustered in at leasttwogenotypes. • Genotype 1 withAsian and Africanisolates • Genotype3 with American and Europeanisolatesfromswine and human UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

  20. 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.

  21. Thank you for your attention! 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 UWI, St. Augustine, Trinidad & Tobago, 29.01.2010.

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