Phylogenetic Analysis of Full-LengthpolGene from Korean Hemophiliacs and Plasma Donors Infected with Korean Subclade B of HIV Type 1

Abstract

There was an outbreak of HIV-1 transmission among 20 out of 122 Korean hemophiliacs from 1990. We assessed the genetic relationships among HIV-1 viruses found in three cash-paid plasma donors whose preseroconversion plasma was used to produce Korean-made clotting factor, 20 hemophiliacs infected with HIV-1 in Korea, three hemophiliacs infected with HIV-1 from clotting factor manufactured outside Korea, and 71 local control patients infected with the Korean subclade of HIV-1 subtype B (KSB). Full-length pol gene sequences (2841 bp) of viruses from frozen stored serum, samples obtained 1-3 years after diagnosis, were amplified by RT-PCR and sequenced by direct DNA sequencing. Phylogenetic and signature pattern analyses were used to investigate the relationships among the sequences. Donors O and P were associated with two clusters, of 8 and 12 hemophiliacs, respectively, which were demarcated from the 71 KSB-infected local control patients and donor R. These data confirm that HIV-1 transmission to 20 hemophiliacs occurred through infusion of Korean-made clotting factor.

Phylogenetic Analysis of the EarliestnefGene from Hemophiliacs and Local Controls in Korea

“…11,12 The use of plasma donors complements the findings of previous studies that lacked plasma donor O sequences. 14 It is suspected that viruses from donors O and P contributed to HIV-1 infections in HPs in Korea.…”

“…10 Recently, we used a similar method to determine the genetic relatedness of the pol and vif genes between plasma donors and hemophiliacs (HPs). 11,12 These studies showed that pol and vif gene analyses are more suitable in cases with significant time lags between primary infection and sampling because these gene regions evolve more slowly than env and are subjected to less individual patient-specific selection pressures. On September 29, 2011, the Supreme Court of Korea ruled that Company X was responsible for the infection with HIV-1 of Korean HPs B after reviewing our studies on pol and vif genes (Case no.…”

“…2008 Da16776). 11,12 At present, cash-paid plasma donation is not permitted by law, and heat inactivation is not included in the original procedure, even though RNA from the Hepatitis A virus was amplified in vials of Factors VIII and IX from 1998 to 1999. 13 …”

“…14 It is suspected that viruses from donors O and P contributed to HIV-1 infections in HPs in Korea. 9,11,12 We performed phylogenetic and signature pattern analyses to the samples to investigate the genetic relationships in the nef sequences among plasma donors, HPs, and local controls (LCs), and compared the results with those of previous analyses that used the pol and vif genes. Although analyses of nef gene sequences are weaker than those of other genes used in previous reports, this nationwide and comprehensive nef gene analysis supports previous findings that two of the four identified HIV-infected plasma donors were the most likely sources of viral infection in the 20 HPs.…”

Molecular Epidemiology of Human Immunodeficiency Virus

“…Phylogenetic analyses were also utilized for epidemiologic evaluation of HIV-infected Korean hemophilia patients [6869], and investigation of HIV-1-transmitted drug resistance (TDR) in Korea [6468707172]. Studies on TDR revealed that the drug resistance mutation was not common among treatment-naïve HIV-infected individuals in Korea, and there was little evidence to suggest that TDR in Korea is related to the clonal spread of drug-resistant strains.…”