Kostrikis L G, Shin S, Ho D D
Aaron Diamond AIDS Research Center, Rockefeller University, New York, New York, USA.
Mol Med. 1998 Jul;4(7):443-53.
Human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) strains can be genetically classified into genetic lineages known as genetic types or subtypes according to phylogenetic analyses of complete or partial nucleotide sequences of their genomes. The genetic classification of HIV-1 and HCV strains has important implications for the development of globally effective vaccines and for the management of patients.
A new method, termed combinatorial DNA melting assay (COMA), allows rapid accessing of comparative genetic information between related DNA sequences, making it possible to rapidly and accurately genotype unknown HIV-1 and HCV strains. COMA is mainly based on the differential melting properties of long DNA heteroduplexes. Combinatorial arrays of DNA heteroduplexes are formed when captured PCR-amplified reference DNA with known nucleotide sequences are combined with solution-phase complementary and antigenically labeled DNA with unknown sequences. Genetic divergence between the known and the unknown sequences is inferred as the experimentally derived melting curves of the two strands of the DNA heteroduplexes increasingly diverge.
COMA was successfully applied to the genetic classification of HIV-1 and HCV strains into phylogenetic lineages or subtypes.
Use of this assay should accelerate current efforts to understand the global molecular epidemiology of HIV-1 and HCV and may extend to the genetic characterization of other genetically diverse infectious pathogens associated with numerous diseases.
根据人类免疫缺陷病毒1型（HIV-1）和丙型肝炎病毒（HCV）基因组完整或部分核苷酸序列的系统发育分析，这些病毒株可在基因上分为称为基因类型或亚型的基因谱系。HIV-1和HCV毒株的基因分类对于开发全球有效的疫苗以及患者管理具有重要意义。
一种称为组合DNA熔解分析（COMA）的新方法，能够快速获取相关DNA序列之间的比较基因信息，从而有可能快速准确地对未知的HIV-1和HCV毒株进行基因分型。COMA主要基于长DNA异源双链体的差异熔解特性。当捕获的具有已知核苷酸序列的PCR扩增参考DNA与具有未知序列的溶液相互补且带有抗原标记的DNA相结合时，就会形成DNA异源双链体的组合阵列。随着DNA异源双链体两条链的实验衍生熔解曲线差异越来越大，可推断已知序列和未知序列之间的基因差异。
COMA已成功应用于将HIV-1和HCV毒株基因分类到系统发育谱系或亚型中。
使用该分析方法应能加快当前了解HIV-1和HCV全球分子流行病学的工作，并可能扩展到对与多种疾病相关的其他基因多样的传染性病原体的基因特征分析。
