Novel viruses and viral dynamics in multiple transfusion recipients

描述（由申请人提供）：寻找新的未知血源性病毒（BBV）的研究通常集中在具有临床症状（发热、肝炎等）的供体或受体的个体血液样本上。血友病、镰状细胞病、地中海贫血、无丙种球蛋白血症、移植受体等患者需要长期输血或血液衍生产品，通常通过汇集20- 60，000名献血者的样本进行。鉴于这一事实，我们假设，基于几百个多次输血接受者（MTR）的无偏高通量测序（UHTS）的研究将能够鉴定几种新的BBV，否则只能通过筛选几千个个体血液样品来发现。多中心血友病队列研究（MHCS）受试者接受了通过汇集数千名个体供体的血浆制备的凝血因子的多次输注。此外，大多数MHCS受试者合并感染HIV，使其极易发生继发性病毒感染;因此，我们预计在一般人群中非常罕见的BBV感染将成为MHCS受试者的常见感染。为了检验这些假设，我们使用UHTS（初步结果）分析了16名MHCS受试者中的两个样本，每个样本间隔>4000天（>10年）。我们在第一个和最后一个时间点样本中分别检测到平均3.4和10.4种不同的病毒。使用物种特异性PCR和克隆测序进行的进化分析表明，不同的BBV物种随时间的不同的遗传多样性，组成和协方差。值得注意的是，除了发现HIV、HCV、HBV、GBVc、TTV和细小病毒B19的高流行率外，我们还发现了几种最近在人类血液样本中发现的从未报道过的病毒和几种新病毒。我们的目标是对人类血清样本中发现的这些新BBV进行遗传表征，使用血清学确认其真实的人类感染，并确定其在不同MTR队列、输血传播病毒研究受试者、健康献血者和其他疾病队列中的感染率。仅在美国，每年约有500万人需要输注人血衍生产品。建议鉴定在人类血液中发现的新病毒、其基因特徴和在不同人口中的流行情况，将有助进行其后的研究，以确定其经输血传播的风险，并制定预防新感染/疾病的策略。

DESCRIPTION (provided by applicant): Studies to find new-unknown blood borne viruses (BBV) generally focus on individual blood samples of donors or recipients with clinical symptoms (fever, hepatitis, etc.). Patients with hemophilia, sickle cell disease, thalassemia, agammaglobulinemia, transplant recipients and others, require chronic transfusion of blood or blood derived products, often made by pooling samples of 20-60,000 blood donors. Given this fact, we hypothesize that an unbiased high- throughput sequencing (UHTS) based study of a few hundred multiple transfusion recipients (MTR) will enable identification of several new BBV that otherwise can only be found by screening several thousands of individual blood samples. Multicenter Hemophilia Cohort Study (MHCS) subjects received multiple transfusions of coagulation factors made by pooling plasma of thousands of individual donors. Moreover, a majority of MHCS subjects are co-infected with HIV, making them highly susceptible to secondary virus infections; therefore we anticipate that the BBVs that are very rare infections in general population will be common infections in MHCS subjects. To test these hypothesis, we used UHTS (preliminary results) to analyze two samples each of 16 MHCS subjects separated by >4000 days (>10 yrs.). We detected an average of 3.4 and 10.4 different viruses in first and last time point samples, respectively. Evolutionary analyses done using the species specific PCR and clonal sequencing indicated distinct genetic diversity, composition and co-variance of different BBV species over time. Noticeably, in addition to finding high prevalence of HIV, HCV, HBV, GBVc, TTV and Parvovirus B19, we found several recently identified viruses never before reported in human blood samples and several new viruses. Our goal is genetic characterization of these new BBV found in human serum samples, confirm their authentic human infection using serology and determine their infection prevalence in different MTR cohorts, Transfusion Transmitted Virus Study subjects, healthy blood donors and other disease cohorts. Approximately 5 million individuals require transfusion of human blood derived product every year in the United States alone. The proposed identification of new viruses found in human blood, their genetic characterization and prevalence in different population will help in conducting subsequent studies to determine their risk of transfusion transmission and in developing strategies to prevent new infections/diseases.