HIV-1耐药性基因型检测法在临床治疗随访中的应用

Application of HIV - 1 genotypic resistance testing in clinical treatment and follow - up

目的 探索HIV-1耐药性基因型检测法在高效抗逆转录病毒疗法（HAART）治疗组中监测HIV-1耐药性病毒株的临床应用。方法 从接受HAART的HIV-1感染者血浆中抽提病毒RNA,采用套式RT-PCR方法扩增HIV-1的PR和RT基因片段,并对扩增片段进行序列测定和分析。结果HIV-1耐药性基因型检测法能够从血浆病毒载量在 1000拷贝/ml以上的样本中得到扩增产物。在 16例接受 HAART的 HIV-1感染者中,发现 1例感染者 DP31的 PR和 RT基因出现了突变,这些突变为L63P、T215F、K219Q和M184V。所有突变均与公开报道的结果一致,并被证明与HIV-1的耐药性有关。另外,DP31在接受HAART前已出现T215F、K219Q的耐药性突变,究其原因,还有待进一步研究。结论HIV-1耐药性基因型检测法能有效地监测接受HAART的HIV-1感染者血浆中耐药性病毒株的存在。该方法提供的结果准确、可靠,并且为临床医生评价HAART效果,合理选择和及时优化药物组合方案提供了可靠的依据。

Objective To explore the feasibility of HIV - 1 genotypic resistance testing and its potential clinical application in HIV- infected patients receiving highly active antiretroviral therapy(HAART) . Methods HIV- 1 viral RNA was extracted from plasma samples of HIV - infected patients receiving HAART and amplified by nested RT - PCR using specific primers. Nucleotide sequences of HIV- 1 protease (PR) and reverse transcriptase(RT) genes were determined by automated DNA sequencer and analyzed using specific biological software. Results RT - PCR products were obtained from the plasma samples of patients whose viral load was as low as 1000 copies/ml.Specific drug- resistant mutations were detected in the PR and RT genes of one (DP31) out of 16 HIV - infected individuals, which were L63P, T215F, K219Q and M184V mutations. In addition, T215F and K219Q mutations were emerged in DP31 before the initiation of HAART, indicuating these drug - resistant viruses might have been transmitted during the initial acquisition of HIV infection.Conclusion HIV- 1 genotypic resistance testing can be used effectively to detect drug resistant viruses in plasma of patient on HAART. The results of genotyping may provide physicians with information for choosing the appropriate antiretroviral drugs and optimizing regimens