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Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” MOLECULAR CHARACTERIZATION OF HAV ISOLATES IN ARGENTINA.

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Presentation on theme: "Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” MOLECULAR CHARACTERIZATION OF HAV ISOLATES IN ARGENTINA."— Presentation transcript:

1 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” MOLECULAR CHARACTERIZATION OF HAV ISOLATES IN ARGENTINA Jorge E. González National Reference Laboratory for Viral Hepatitis. INEI-ANLIS “Dr. Carlos G. Malbrán”. Buenos Aires. Argentina

2 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” HAV INFECTION IN ARGENTINA (antiHAV Prevalence) 1) Healthy children under 10 years old n=3 699 HAV INFECTION IN ARGENTINA (antiHAV Prevalence) 1) Healthy children under 10 years old n= ) Blood donors (Greater BA) n=1156 Acta Gastr Latin 27 (1997 ) unpublished antiHAV 1 dot = 100 residents/km 2

3 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” LIVER TRASPLANT at Garrahan hospital The main pediatric hospital in Argentina LIVER TRASPLANT at Garrahan hospital The main pediatric hospital in Argentina Cuarterolo M et al. Liver trasplant. Long follow up. Medicina Infantil 2002; IX: n = 143 X age: 5.4 yr Range: 0.6 – 22 yr 53%  Period:11/92 – 11/01

4 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” to characterize HAV strains circulating in Buenos Aires (BA) area and in the rest of the country. Objective

5 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN”LOCALITYProvinceDATESRESIDENTS Nº OF CASES AGE DISTRIBUTION Barreal San Juan May yr: 14% 5-9 yr: 42% yr: 23% > 14 yr : 5% Undated : 16% Maquinchao Rio Negro August yr : 15% 5-9 yr : 67% yr: 13% yr: 4% >20 yr : 2% Gan Chubut January yr: 14% 5-8 yr: 63% 9-11 yr: 23% HAV outbreaks 1 dot = 100 residents/km 2

6 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” HAV sporadic cases LOCALITYProvince San Justo Greater B A Bernal City of Buenos Aires DATESDec 97-Sep 98 Oct 01–May 02 May 2002 June 1999 Nº OF CASES AGEDISTRIB yr 7yr and 8 yr 41 yr22 yr

7 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” Methods ELISA for antiHAV-IgM (HAVAB-M EIA, Abbott USA ) in-house RT nested PCR using primers targeting VP1-2A region. Sequencing using a Big Dye DNA sequencing kit (Perkin Elmer) on a 373 DNA Secuencer apparatus ( Perkin Elmer) Phylogenetics analisys (Bioedit, Clustal, Philips package).

8 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” RESULTS (I) ALIGNMENT OF PARCIAL NUCLEOTIDE SEQUENCES OF HAV ISOLATES (168 BASES IN THE VP1-2A JUNCTION)

9 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” ALIGNMENTS OF THE AMINO ACID FROM THE VP1-2A JUNCTION RESULTS (II)

10 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” RESULT (III) 0.1 SFL CF53 AGM CY145 JM55 KPH H132 PA21 GA76 HM175 ARG6 Arg5.3 & 5.10 (San Justo, Mar 98 / Dic 97) (2) Arg4.10 (San Justo, Oct 01) GBM Arg1.5 ( Maquinchao, Aug 97) ARG23 Arg1.10 (San Justo, Mar 98) ARG1 Arg3.17 ( Bernal, May 02) Arg5.4 (San Justo, Sep 98) Arg6.12 & 0.2 (Gan Gan, Jan 02) (2) ARG4 Arg4.7 & 4.8 & 4.9 (Barreal, May 02) (3) Arg2.22 & 2.19 (San Justo, May 02; BsAs, Jun 99) (2) Arg1.9 (San Justo, Mar 98) Arg1.7 (San Justo, Dic 97) Arg5.5 (San Justo, Apr 98) Arg5.7 (San Justo, Jun 98) VII II IB IA VI IV V IIIB IIIA III I 74 PHYLOGENETIC ANALYSIS OF THE VP1-2A REGION OF HAV ISOLATES (NEIGHBOR-JOINING)

11 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN”CONCLUSIONSCONCLUSIONS All the isolates reported here were sub-genotype IA, so this is still the only one reported in Argentina to date. There were no differences between outbreak and sporadic involved strains and hence, between high (BA area, SJ, Be) and low (Ga, Ba, Ma) population density areas, thus confirming the high endemicity of this infection. The high genetic diversity in this small fragment did not allow us to identify a pattern of circulation. This study will be very important as reference data in the characterizacion of the strain/s producing fulminant cases, since this severe clinical presentation was not the case in any of the infections studied in this series.

12 Servicio Hepatitis y Gastroenteritis Laboratorio Nacional de Referencia INEI-ANLIS “Dr C. G. MALBRAN” LNR Dpto Virología INEI Jorge E Gonzalez Sara N. Vladimirsky Maria S. Munné Lucio O. Otegui Raul E. Castro Leonardo S. Brajterman Sonia S. Soto H. Niños San Justo Maria C. Cañero Velasco Acknowledgments Acknowledgments -Marcela Nobile Prov. Rio Negro -Maria Jofre Prov. San Juan -Eduardo Fernandez Prov. Chubut -Octavio Fusco INP “Fatala Chaben” -Jorge Quarleri Facultad Medicina UBA A.N.L.I.S. “Dr Carlos G. Malbran”


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