Novel viruses and viral dynamics in multiple transfusion recipients

描述（由申请人提供）：寻找新未知血源性病毒（BBV）的研究一般集中在有临床症状（发热、肝炎等）的献血者或受体的个体血液样本上。血友病、镰状细胞病、地中海贫血、无球蛋白血症患者、移植受者和其他患者需要长期输血或血液制品，通常需要汇集20-60 000名献血者的样本。鉴于这一事实，我们假设对数百名多次输血受者（MTR）进行无偏倚的高通量测序（UHTS）研究将能够识别几种新的BBV，否则只能通过筛选数千个个体血液样本来发现。多中心血友病队列研究（MHCS）的受试者接受多次输血凝血因子，这些凝血因子是由数千名个体献血者的血浆汇集而成的。此外，大多数MHCS受试者同时感染艾滋病毒，使他们极易继发病毒感染；因此，我们预计在一般人群中非常罕见的bbv感染将在MHCS受试者中常见。为了验证这些假设，我们使用UHTS（初步结果）对16名MHCS受试者中的两个样本进行了分析，每个样本相隔4000天（10年）。我们在第一个和最后一个时间点样本中分别平均检测到3.4和10.4种不同的病毒。利用物种特异性PCR和克隆测序进行的进化分析表明，随着时间的推移，不同BBV物种具有不同的遗传多样性、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，我们还发现了一些最近在人类血液样本中从未报道过的病毒和一些新病毒。我们的目标是在人类血清样本中发现这些新的BBV的遗传特征，用血清学证实它们的真实人类感染，并确定它们在不同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流行率。仅在美国，每年大约有500万人需要输血人血制品。拟议确定人类血液中发现的新病毒、它们的遗传特征和在不同人群中的流行情况，将有助于开展后续研究，以确定其输血传播的风险，并有助于制定预防新感染/疾病的战略。

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.