Next-generation sequencing technology:A technology review and future perspective

As one of the most powerful tools in biomedical research,DNA sequencing not only has been improving its productivity at an exponential growth rate but has also been evolving into a new layout of technological territories toward engineering and physical disciplines over the past three decades.In this technical review,we look into technical characteristics of the next-generation sequencers and provide insights into their future development and applications.We envisage that some of the emerging platforms are capable of supporting the USD1000 genome and USD100 genome goals if given a few years for technical maturation.We also suggest that scientists from China should play an active role in this campaign that will have a profound impact on both scientific research and societal healthcare systems.

Application of high-throughput sequencing technology in HIV drug resistance detection

Drug resistance via drug-resistant mutations in the human immunodeficiency virus(HIV)genome is the primary cause of antiviral therapy failure.Consequently,HIV drug resistance genotyping has become a critical approach in HIV prevention and control.Compared to the Sanger sequencing technology,high-throughput sequencing(HTS)technology has superior sensitivity and timeliness,with strong detection capabilities for low-frequency mutations.With the continued advancement of HTS technologies,their prominence in HIV drug resistance detection techniques has increased accordingly.This article will review the latest developments in HTS technology and its applications in HIV drug resistance testing.

Novel applications of next-generation sequencing in breast cancer research

With the rapid development of medicine,the studies of genes have become increasingly concerned by more people and being the contend of a great of researches.The next generation sequencing with its own advantages has been widely used in gene research nowadays.It has almost replaced the traditional sequencing methods(such as Sanger sequencing method),and played an important role in a variety of complex disease researches,including breast cancer.The next generation sequencing technology has the advantages of high speed,high throughput and high accuracy.It has been widely used in various cancers(such as prostate cancer,lung cancer,pancreatic cancer,liver cancer,etc.),especially in breast cancer.Moreover,the use of the next generation sequencing technology to make DNA sequence analysis and risk prediction has made a great contribution to the research of breast cancer.We will focus on the application of whole genome sequencing,exon sequencing and targeted gene sequencing in breast cancer gene research.

Variations in microbial community during nitrogen removal by in situ oxygen-enhanced indigenous nitrogen-removal bacteria

In this study, the enclosure system exhibited perfect nitrogen removal performance with in situ oxygen-enhanced indigenous aerobic denitrifying bacteria in an enclosure experiment. We explored changes in the microbial community during the nitrogen removal process using the MiSeq high-throughput sequencing technology. The results revealed a total of 7974 and 33653 operational taxonomic units(OTUs) for water and sediment systems, respectively, with 97% similarity. The OTUs were found to be affiliated with eight main phyla(Proteobacteria, Actinobacteria, Cyanobacteria, Bacteroidetes, Planctomycetes, Chloroflexi, Firmicutes, and Actinobacteria). The diversity of the enhanced system was found to be higher than that of the control system. Principal component analysis(PCA) revealed that significant spatial and temporal differences were exhibited in the microbial community during nitrogen removal in the enclosure experiment. Redundancy analysis(RDA)indicated that physical parameters(temperature, dissolved oxygen, and pH), nitrogen(total nitrogen and nitrate), functional genes(nirK and nirS), and dissolved organic carbon(DOC) were the most important factors affecting bacterial community function and composition. Lastly, the results suggested that the variation in the microbial community could be analyzed through the MiSeq high-throughput sequencing technology,which may provide technical support for future field tests.

Microbial diversity in the sediments of the southern Mariana Trench

Microbial diversity in the abyssal sediments beneath the seafloor of 30,94,and 151cm near the southern end of the Mariana Trench was analyzed in the Illumina HiSeq 2500 platform.Results show that the microbial populations were dominated by bacteria but merely no archaea were identifi ed at the three depths.In the bacterial community,Proteobacteria and Firmicutes dominated the total taxon tags,followed by Bacteroidetes,Actinobacteria,Planctomycetes,Cyanobacteria,and Chloroflexi,which together account for over 99%of the total population.Similar to that in the seawater in the trench,the operational taxonomic units(OTUs)belonging to Gammaproteobacteria from the sediment samples showed high abundance.However,common bacterial OTUs in the water of the trench including Nitrospirae and Marinimicrobia were hardly found in the sediments from the southern Mariana Trench or the hadal region.Therefore,this study documented for the first time the compositions of microbial diversity in the trench sediments,revealed the difference in microbial diversity in water and sediment of the trench and will enrich the knowledge on the microbial diversity in the abyssal areas.

Effect of a compound starter cultures inoculation on bacterial profile and biogenic amine accumulation in Chinese Sichuan sausages

Effects of a starter culture composed of Lactobacillus sakei,Pediococcus pentosaceus,Staphylococcus xylosus and S.carnosus at the ratios(m/m)of 1:1:1:1 on bacterial profile and biogenic amine accumulation in Chinese Sichuan sausages were demonstrated.In starter culture-inoculated sausages,Lactobacillus spp.,Pediococcus spp.and Staphylococcus spp.were predominant bacterial genus and their relative abundances ranged from 70.14%to 25.98%,57.66%to 14.08%,and 15.71%to 13.40%during fermentation,respectively.Accordingly,Lactobacillus spp.and Weissella spp.were predominant bacterial genus and their relative abundances ranged from 63.14%to 24.70%and 49.40%to 11.96%during spontaneous fermentation,respectively.Furthermore,the abundances of undesirable microorganisms such as Yersinia spp.,Enterobacter spp.,Acietobacter spp.and Psychrobacter spp.were lower than that of the control.The levels of histamine,putrescine,tyramine and cadaverine in Chinese Sichuan sausages with starter cultures inoculation were significantly lower(P<0.05)than that of the control,and were decreased by 83.09%,69.38%,51.87%and 57.20%,respectively,at the end of the ripening.These results revealed that the starter cultures inoculation was a better alternative for microbial quality improvement and biogenic amine reduction of Chinese Sichuan sausages with good sensory attributes.

Tmap:An Integrative Framework for Studying Population-scale Microbiome

With the advent of next generation sequencing technologies,conventional data analysis methods are becoming inadequate for the analysis of highly complex and voluminous microbiome data.In particular,nonlinear associations hidden in a high-dimensional microbiome dataset are hard to capture using common linear-based methods.

新一代测序技术的研究进展

大规模DNA测序技术是揭秘人类和其它生物遗传密码的重要技术,在分子生物学和基础医学领域有广泛应用。第二代测序技术的出现使DNA测序的通量大幅提高,测序的成本大幅下降,原来只有在大型测序中心才能完成的测序任务现在已经可以在更多的实验室展开。但是,早期的第二代测序技术仍然存在诸如文库构建过程复杂、测序成本依然较高等缺点。为了克服上述缺点,近三年发展了几种新的第二代和第三代测序技术,这些技术不仅继承了早期第二代测序技术通量高的优点,而且在文库构建等方面取得了重要突破,进一步简化了测序操作,降低了测序成本,缩短了测序时间。本文就几种最新的大规模测序技术的原理、特点与发展趋势进行简要介绍。