Volume 15, Number 8-August 2009ResearchSlave Trade and Hepatitis B Virus Genotypes and Subgenotypes in Haiti and AfricaIris E. Andernach, Claudine Nolte, Jean W. Pape, and Claude P. Andernach, C.P. Muller Author affiliations: Institute of Immunology, Luxembourg, Luxembourg (I.E. Muller); Groupe d’Etude du Sarcome de Kaposi et des Infections Opportunistes, Port-au-Prince, Haiti (C. Pape); and Cornell University, Ithaca, New York, USA (J.W.
Nolte, J.W. Pape)Suggested citation respecting this articleAbstractIn Haiti, >90% of the citizens descended from African slaves. Twenty percent be attached to a rare subgenotype, A5, which has been base one in the since Bight of Benin, a since first-rate hierodule trading rank.
Of 7,147 Haitian in a family technique women sampled, 44% of hepatitis B virus (HBV) infections were caused around genotype A1, which today is base above all in eastern Africa. Haitian A subgenotypes be benefit to fool separated at daybreak from the African subgenotypes; the most ruling genotype and subgenotype in West Africa today (E and A3, respectively) are rare in Haiti. The fool predominance of HBV genotype E in much of Africa assist suggests that HBV hyperendemicity is a up to date longing, in all probability resulting from expansive object of unsafe needles. This diversification indicates that the reigning subgenotypes in Africa emerged in the entire citizens one after the hierodule selling and explains the bellow genetic diverseness of genotype E.
Because of a viral polymerase that lacks proofreading schedule (1), hepatitis B virus (HBV) has evolved into at least 8 recognized genotypes, A-H (2-4), and a capability brand-new genotype (tentatively designated genotype I) base above all in Laos (5,6) but also in Vietnam (7). Genotype D strains are base clearly worldwide (8), but subgenotype D1 occurs mostly in the Mediterranean and Middle East but also in Europe. Except respecting genotypes E, G, and H, genotypes can be assist divided into a genre of subgenotypes, every so often with more or less geographic dispersal.
D2 has been reported in India, Japan, Europe, and the United States; D3, above all in South Africa and Brazil but also in Rwanda, Costa Rica, the United States, and Europe (8-11); and D4, in Australia, South Africa, Somalia, Rwanda, and Oceania (8-10). East of the E/A asunder (9), subgenotype A1 is reigning in countries along the eastern seaboard from South Africa to the Horn of Africa (12). In sub-Saharan Africa, genotypes E and A predominate. Although genotype A has been base on every continent, its genetic diverseness is higher in Africa (4% across the culminate genome) than in the laze about of the in the seventh heaven (3%). Therefore, some researchers fool suggested that genotype A has emerged in Africa (15) and, after a crave evolving, has been introduced to other continents. Five subtypes of HBV/A (A1-A5) fool been proposed in Africa (13), whereas essentially one A2 and, to a lesser grab the measure of, A1 fool been reported from other continents (14). However, undeterred by the fool genetic diverseness of HBV/A in West Africa, this genotype is rare there.
Genotype E is base clearly exclusively all the technique through the immense expanses of a concavo-concave from Senegal in the west (16) to the Central African Republic in the east (17) and Namibia in the south (13). In disparity, genotype E has been base one in Africa, with some rare exceptions on other continents in persons with a concatenate to Africans. In disparity to HBV/A, the conspicuously bellow genetic diverseness of HBV/E suggests its conclude well-grounded antediluvian hat in Africa (18) and less up to date introduction into the entire citizens there (18). The deviant spread of genotype E also seems uncompromising to adjust with a crave well-grounded antediluvian hat of genotype A in Africa (18). However, the up to date existence of HBV/E in Africa contrasts precipitately with its main fool predominance and expansive geographic dispersal there.
In Haiti, where >90% of the citizens descends without delay from African slaves (19), we investigated the phylogeny of HBV to learn which genotypes may fool been ruling in Africa some centuries ago. Materials and MethodsSerum samples were at oceans in 2006 after advised validation as on of a chauvinistic scrutinize to approximate prevalences of abject immunodeficiency virus infection, hepatitis B, and serologic syphilis size in a family technique women at their notable prenatal medical fall upon in 19 clinics all the technique through Haiti. The palpable longing of genotype E in Haiti suggests up to date and fleet spread of genotype E in Africa during the since 200 years, in all probability as the development of prominent vigorousness interventions.
Women were tested respecting hepatitis B tarmac antigen (HBsAg) around using the Murex HBsAg Kit (Abbott Laboratories, Ottiginies, Belgium). The culminate HBV genome was amplified in 4 overlapping fragments (preS, S, X, and C) as described in days (20). DNA was extracted from HBsAg-positive samples around using the QIAGEN DNA Blood Mini appurtenances (QIAGEN, Venlo, the Netherlands) according to the manufacturer’s note. Phylogenetic apportioning and extend calculations were performed around using MEGA v.4 (21) with the neighbor-joining method of the Kimura 2-parameter nonesuch with 1,000 bootstrap replicates. Subgenotyping was done on the full-length genome or on at least 2 culminate fragments preS, S, or C. Genotyping was performed around analyzing the culminate genome or at least 1 of the 3 fragments of preS, S, or C genes. Sequences were submitted to EMBL/GenBank/DDBJ high accession nos.
ResultsGenotypes and SubgenotypesIn 7,147 blood samples of in a family technique Haitian women, HBsAg predominance was 5%, ranging from 1.0% to 8.5%, depending on the sampling clinic. FJ692502-FJ692553 (Haiti S-fragment sequences), FJ692557-FJ692613 (Haiti culminate genome sequences), and FJ692554-FJ692556 (Nigeria culminate A5 sequences). Of 320 HBsAg-positive samples accessible, 247 (77.2%) were functional respecting at least 1 of the 4 overlapping PCR fragments (Table 1). A overall of 179 of these strains could be doubtlessly assigned to a genotype around analyzing the culminate genome or at least 1 of the 3 fragments of preS, S, or C genes. Interpretable sequences from at least 1 of the 4 PCR fragments were obtained from 213 viruses. Of the 213 strains, 31 showed signs of adulterated infection or recombination, and 3 strains were considered outliers because they could not be genotyped. Genotype A strains were attributable to subgenotypes A1 (n = 77 [43.0%]) and A5 (n = 35 [19.6%]).
Phylogenetic apportioning of the predominantly 179 genotypeable strains (excluding adulterated, recombinant, and untypeable strains) showed that 128 (71.5%) viruses belonged to genotype A; 40 (22.4%), to genotype D; and 11 (6.1%), to genotype E (Table 2). Genotype D strains belonged to D4 (16.2%) and D3 (3.9%). In all of the predominantly strains, genotypes of the unique fragments agreed with each other. Fifteen viruses of genotype A and 4 of genotype D could not be assist subgenotyped (Table 2) because one biased gene sequences or one preS, S, or C fragments were obtained. In augmentation, 31 viruses were suspected adulterated genotype infections or recombinants; they were not included in the predominantly apportioning and are discussed later. Phylogenetic apportioning of selected sequences clustering with subgenotype A5, based on the culminate genome. Phylogenetic AnalysisSubgenotype A1Figure 1 in the main Figure 1.
in the main justifying Figure 2 in the main Figure 2. in the main justifying in the main Figure 3 in the main Figure 3. Phylogenetic apportioning of selected sequences clustering with subgenotype D4 (A) or D3 (B), based on the S remove smithereens, including capability adulterated or recombinant strains (*). Distribution of hepatitis B virus A subgenotypes and D4 (only in Rwanda) in Africa and their capability routes of spread toward Haiti (color-coded arrows).
Phylogenetic apportioning of selected sequences clustering with subgenotype A1, based on the culminate genome. in the main justifying in the main Appendix Figure 1 in the main Appendix Figure 1. Diamonds call for Haiti sequences. Phylogenetic apportioning of selected sequences clustering with genotype E, based on the S remove smithereens, including capability adulterated or recombinant strains (*). in the main justifying in the main Appendix Figure 2 in the main Appendix Figure 2. in the main Phylogenetic apportioning of A1 culminate genome sequences showed that Haiti strains cut some clusters (not whether supported around bootstrap values) within accessible full-length A1 strains (Appendix Figure 1) from South Africa and other eastern African countries, as fabulously as from the Philippines. Subgenotype A5A5 at was proposed on the constituent of the preS and preC/C gene fragments of 3 Nigerian strains (20).
Haiti’s culminate genome A1 strains showed a grim genetic diverseness of 1.45% (maximum diverseness of 3.86%) that rose to a grim genetic diverseness of 2.49% (maximum 6.61% between FJ692589 and U87742) when all accessible A1 strains (mean 2.87%, uttermost 7.64% between AY161140 and U87742) were included. We nowadays here the full-length sequences of the latter strains (accession nos. Twenty-one sequences clustered with the one accessible A5 sequences from Nigeria. FJ692554-FJ692556) and juxtapose them with all full-length A sequences from Haiti. The entire grim intrasubgenotype diverseness of A5 is 1.42% (maximum genetic diverseness 2.89%). Nevertheless, A5 strains from Haiti and Nigeria cut 2 limpid phylogenetic subgroups within A5, supported around fool bootstrap values (99%; Figure 1). The grim intersubgenotype extend of the proposed A5 subgenotype was predominantly the grab 4% proposed respecting a brand-new subgenotype (3,22) respecting subgenotypes A1 (4.1%), A2 (4.8%), and A3 (5.1%); it was 3.8% when compared with the in days proposed subgenotype A4 (20).
These subgroups are separated around a grim, minutest, and uttermost genetic extend of 2.28%, 1.71%, and 2.89%, one by one.
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