USING MICROSATELLITES TO ANALYZE CYTOMEGALOVIRUS STRAINS

使用微卫星分析巨细胞病毒株

• 批准号: 2672998
• 负责人: CHRISTOPHER J WILLS
• 金额: $ 7.58万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2672998
• 关键词: cytomegalovirus; drug resistance; foscarnet; ganciclovir; genetic markers; genetic polymorphism; genetic strain; genome; genotype; nucleic acid sequence; polymerase chain reaction; serotyping; virulence; virus infection mechanism

DESCRIPTION (adapted from applicant's abstract) The ability to distinguish morphologically identical strains of pathogens is critical to proper diagnosis and treatment of opportunistic infections such as cytomegalovirus in AIDS patients. The investigators propose to use microsatellites to investigate important questions of transmission, virulence and drug resistance in the herpes-like virus, cytomegalovirus. While relations between genotype and virulence have been detected for other genetic markers, microsatellites may provide a large number of new markers to aid in the understanding of CMV virulence. Microsatellite sequences are short arrays of direct repeats which accumulate length mutations due to frequent replication slippage errors. The investigators have found that these loci occur frequently enough even in small genomes (8000 bp to 50 Mbp) to make them a very useful, and largely untapped, source of genetic markers in microorganisms. DNA sequencing of such loci often detects alleles that have the same length but a different sequence, allowing them to distinguish among various strains with high certainty. In a preliminary survey, four such repeats have been located in the cytomegalovirus genome, and the investigators have also identified suitable loci for study in Cryptosporidium parvum, Pneumocystis, Cryptococcus, and Mycobacterium. The development of microsatellite markers for such organisms represents a novel approach to the characterization of opportunistic infections, and may allow them to follow outbreaks in significant detail. The applicants intend to use this approach to examine relationships among genotype, virulence and drug resistance in cytomegalovirus. This information may aid in therapy and prevention. The investigators' data from C. albicans show the feasibility of microsatellite typing for distinguishing among strains and for studying the evolution of resistance to fungistatic agents in strains isolated from HIV-positive individuals. Similar approaches will be taken to the study of strains of cytomegalovirus, which have a great range of effects on host cells and which readily give rise to mutants resistant to antiviral agents.

描述（改编自申请人摘要）区分能力 形态上相同的病原体菌株对正确的 诊断和治疗机会性感染，如巨细胞病毒 在艾滋病患者中。 研究人员建议使用微型卫星， 调查传播、毒力和药物等重要问题 疱疹样病毒巨细胞病毒的抗药性。 虽然基因型和毒力之间的关系已被发现为其他 遗传标记、微卫星可能提供大量新的标记 以帮助理解CMV的毒力。 微卫星序列 短的同向重复序列阵列，其积累由于以下原因而导致的长度突变： 频繁的复制滑动错误。 调查人员发现， 这些位点即使在小基因组（8000 bp至50 Mbp）中也足够频繁地出现 使其成为一个非常有用的，而且在很大程度上尚未开发的遗传标记来源 在微生物中。 这些基因座的DNA测序通常检测到 有相同的长度，但不同的序列，让他们区分 在不同的菌株中有很高的确定性。 在初步调查中， 这样的重复序列已经位于巨细胞病毒基因组中， 研究人员还确定了适合研究的基因座， 隐孢子虫、肺孢子虫、隐球菌和分枝杆菌。 的 微卫星标记的发展，这些生物代表了一种新的 机会性感染的表征方法，并可能允许 他们对疫情进行了详细的跟踪。 申请人旨在 使用这种方法来检查基因型，毒力和 巨细胞病毒的耐药性 这些信息可能有助于治疗， 预防 研究者从C.白色念珠菌显示了 微卫星分型用于区分菌株和研究 菌株对抑真菌剂的耐药性演变 艾滋病毒阳性者。 将采取类似的方法研究 巨细胞病毒株，对宿主有广泛的影响 细胞，并且容易产生抗病毒剂的突变体。