Clarke John R
Department of Genitourinary Medicine, Division of Medicine, Imperial College of Science, Technology and Medicine, St. Mary's Campus, Paddington, London W2 1PG, UK.
Expert Rev Mol Diagn. 2002 May;2(3):233-9. doi: 10.1586/14737159.2.3.233.
The development of molecular techniques that access viral load and the development of genotypic resistance have revolutionized the treatment of HIV disease. Commercially available viral load assays use a number of different approaches from reverse transcriptase PCR to amplification of branched chain DNA. The drawbacks of the assay are that there is no international standard that allows comparison of viral load between assays and the diversity of different clades of HIV results in the under or the nondetection of some patients samples. New real-time PCR assays are under development, including LightCycler- and TaqMan-based tests. The development of sequence-based genotyping assays for the detection of mutations associated with the development of the resistance to the 17 licensed drugs targeted against the pol gene of HIV have added to the improvements in patient management. However, next-generation assays must extend detection to include the gp41 fusion region and the integrase region of the genome as compounds directed against these targets move from clinical trails into licensed drugs. Also, genotypic assays must improve detection of minor species and detection of sequences from patients with low viral load number. Real-time sequence based-diagnostics remains a realistic target within the next 5 years.
能够检测病毒载量的分子技术的发展以及基因型耐药检测技术的发展，彻底改变了HIV疾病的治疗方式。市售的病毒载量检测方法采用了多种不同的技术，从逆转录酶PCR到分支链DNA扩增。这些检测方法的缺点在于，没有国际标准来实现不同检测方法之间病毒载量的比较，而且HIV不同进化枝的多样性会导致一些患者样本检测不到或检测结果偏低。新型实时PCR检测方法正在研发中，包括基于LightCycler和TaqMan的检测。用于检测与针对HIV pol基因的17种已获许可药物耐药性相关突变的基于序列的基因分型检测方法的发展，进一步改善了患者管理。然而，随着针对这些靶点的化合物从临床试验进入获批药物阶段，下一代检测方法必须将检测范围扩大到包括基因组的gp41融合区和整合酶区。此外，基因分型检测必须提高对次要毒株的检测能力以及对病毒载量低的患者序列的检测能力。基于实时序列的诊断在未来5年内仍是一个切实可行的目标。
