高效抗逆转录病毒治疗的HIV/HBV重叠感染者耐药HBV毒株C区和P区基因序列分析

Sequence detection of HBV-DNA P and C region in HIV/HBV superinfection subjects with drug resistance to HAART

目的 :进一步研究乙肝病毒 (HBV)耐高效抗逆转录疗法 (HAART)的耐药机制。方法 :用 HBV- DNA荧光定量试剂盒 ,定量检测 36例 HAART治疗的 HIV/ HBV重叠感染者中 HBV- DNA,用 HBV- DNA C区和 P区引物常规扩增 HBV- DNA C区和 P区产物 ,PCR产物纯化后测序 ,用 BL AST软件分析序列 ,与国际 HBV基因库对比。结果 :36例 HAART治疗的 HIV/ HBV重叠感染者中 ,HBV- DNA阳性 4例 (11.1% ) ,血清中 HBV- DNA拷贝数 3例在 10 4copies/ ml级 ,1例在 10 7copies/ ml级。对 1例 HBV- DNA在 10 7copies/ ml级标本进行 HBVDNA C区和 P区序列测定 ,用 BL AST软件与国际基因库对比 ,结果 ,C区 nt 2 4 12位 T→ C,nt 2 4 13位 T→ C,P区 nt 74 1位A→ G(YMDD→ YVDD)。结论 :HAART治疗的 HIV/ HBV重叠感染者 ,HBV耐药可能与 HBV- DNA C区和 P区联合变异有关。

Objective: To further study the resistance of HBV to high activity antiretrovirus therapy(HAART). Methods: HBV-DNA was quantitatively detected by real-time PCR in 36 HIV/HBV superinfection subjects, C region and P region HBV-DNA in high copies HBV-DNA subjects were detected by routine PCR, PCR products were purified and sequenced and compared with the HBV international Genbank using BLSAT softy ware. Results: HBV-DNA was positive in 4 of 36 patients(11.1%) and another 3 had low copies(<10 4copies/ml), one had a high HBV-DNA copies(10 7copies/ml). It's HBV-DNA C region sequence had mutation on 2 sites (nt 2412 T/C; nt 2413 T/C)and 1 mutation P region (nt 741 A/G, also YMDD/YVDD) compared with HBV international Genbak reference sequence. Conclusion: The HBV resistance to HAART may be related with multiple genetic mutations in the C and P regain of HBV-DNA.