Viral Nucleic Acids in Live-Attenuated Vaccines: Detection of Minority Variants and an Adventitious Virus
Viral Nucleic Acids in Live-Attenuated Vaccines: Detection of Minority Variants and an Adventitious Virus
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DOI：
10.1128/jvi.02690-09
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发表时间：
2010-06-01
影响因子：
5.4
通讯作者：
Delwart, Eric L.
中科院分区：
文献类型：
--
作者：
Victoria, Joseph G.;Wang, Chunlin;Delwart, Eric L.
宏基因组学和泛微生物微阵列被用来检查八个减毒活病毒疫苗。对三价口服脊髓灰质炎病毒（OPV）、风疹、麻疹、黄热病、水痘-带状疱疹、多价麻疹/腮腺炎/风疹和两种轮状病毒活疫苗中的病毒核酸进行部分纯化、随机扩增和焦磷酸测序。产生了超过50万个序列读段，覆盖了20%至99%的减毒病毒基因组，深度达到每个核苷酸8，000个读段。在口服脊髓灰质炎疫苗、腮腺炎病毒和水痘-带状疱疹病毒中检测到了与疫苗株相关的突变和少数变异，这些变异对减毒的影响未知。证实了从生产禽类和灵长类动物细胞中预期检测到内源性逆转录病毒序列。禽白血病病毒（ALV）以前被证明对人类没有感染性，但在病毒颗粒中以RNA形式存在，而猿猴逆转录病毒（SRV）则以遗传缺陷DNA形式存在。Rotarix是一种口服轮状病毒疫苗，含有猪圆环病毒-1（PCV 1），这是一种高度流行的非致病性猪病毒，尚未证明在人类中具有传染性。将疫苗核酸与泛微生物微阵列杂交证实了内源性逆转录病毒和PCV 1核酸的存在。因此，深度测序和微阵列可以检测减毒病毒序列变化、少数变异和外源病毒，并有助于维持减毒活病毒疫苗的现有安全记录。
Metagenomics and a panmicrobial microarray were used to examine eight live-attenuated viral vaccines. Viral nucleic acids in trivalent oral poliovirus (OPV), rubella, measles, yellow fever, varicella-zoster, multivalent measles/mumps/rubella, and two rotavirus live vaccines were partially purified, randomly amplified, and pyrosequenced. Over half a million sequence reads were generated covering from 20 to 99% of the attenuated viral genomes at depths reaching up to 8,000 reads per nucleotides. Mutations and minority variants, relative to vaccine strains, not known to affect attenuation were detected in OPV, mumps virus, and varicella-zoster virus. The anticipated detection of endogenous retroviral sequences from the producer avian and primate cells was confirmed. Avian leukosis virus (ALV), previously shown to be noninfectious for humans, was present as RNA in viral particles, while simian retrovirus (SRV) was present as genetically defective DNA. Rotarix, an orally administered rotavirus vaccine, contained porcine circovirus-1 (PCV1), a highly prevalent nonpathogenic pig virus, which has not been shown to be infectious in humans. Hybridization of vaccine nucleic acids to a panmicrobial microarray confirmed the presence of endogenous retroviral and PCV1 nucleic acids. Deep sequencing and microarrays can therefore detect attenuated virus sequence changes, minority variants, and adventitious viruses and help maintain the current safety record of live-attenuated viral vaccines.