High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia
Xiao-Jun Yang,1,* Yan-Bo Wang,2,3,* Zhi-Wei Zhou,4,* Guo-Wei Wang,2 Xiao-Hong Wang,1 Qing-Fu Liu,1 Shu-Feng Zhou,4 Zhen-Hai Wang2,3 1Department of Intensive Care Unit, 2Neurology Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People’s Republic of China; 3Key La...
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| Format: | Article |
| Language: | English |
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Dove Medical Press
2015-08-01
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| Series: | Drug Design, Development and Therapy |
| Online Access: | http://www.dovepress.com/1high-throughput-sequencing-of-16s-rdna-amplicons-characterizes-b-peer-reviewed-article-DDDT |
| _version_ | 1818065016384913408 |
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| author | Yang XJ Wang YB Zhou ZW Wang GW Wang XH Liu QF Zhou SF Wang ZH |
| author_facet | Yang XJ Wang YB Zhou ZW Wang GW Wang XH Liu QF Zhou SF Wang ZH |
| author_sort | Yang XJ |
| collection | DOAJ |
| description | Xiao-Jun Yang,1,* Yan-Bo Wang,2,3,* Zhi-Wei Zhou,4,* Guo-Wei Wang,2 Xiao-Hong Wang,1 Qing-Fu Liu,1 Shu-Feng Zhou,4 Zhen-Hai Wang2,3 1Department of Intensive Care Unit, 2Neurology Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People’s Republic of China; 3Key Laboratory of Brain Diseases of Ningxia, Yinchuan, Ningxia, People’s Republic of China; 4Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA *These authors contributed equally to this work Abstract: Ventilator-associated pneumonia (VAP) is a life-threatening disease that is associated with high rates of morbidity and likely mortality, placing a heavy burden on an individual and society. Currently available diagnostic and therapeutic approaches for VAP treatment are limited, and the prognosis of VAP is poor. The present study aimed to reveal and discriminate the identification of the full spectrum of the pathogens in patients with VAP using high-throughput sequencing approach and analyze the species richness and complexity via alpha and beta diversity analysis. The bronchoalveolar lavage fluid samples were collected from 27 patients with VAP in intensive care unit. The polymerase chain reaction products of the hypervariable regions of 16S rDNA gene in these 27 samples of VAP were sequenced using the 454 GS FLX system. A total of 103,856 pyrosequencing reads and 638 operational taxonomic units were obtained from these 27 samples. There were four dominant phyla, including Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. There were 90 different genera, of which 12 genera occurred in over ten different samples. The top five dominant genera were Streptococcus, Acinetobacter, Limnohabitans, Neisseria, and Corynebacterium, and the most widely distributed genera were Streptococcus, Limnohabitans, and Acinetobacter in these 27 samples. Of note, the mixed profile of causative pathogens was observed. Taken together, the results show that the high-throughput sequencing approach facilitates the characterization of the pathogens in bronchoalveolar lavage fluid samples and the determination of the profile for bacteria in the bronchoalveolar lavage fluid samples of the patients with VAP. This study can provide useful information of pathogens in VAP and assist clinicians to make rational and effective therapeutic decisions. Keywords: bioinformatics, drug resistance, OTU, PCR, DNA sequencing |
| first_indexed | 2024-12-10T14:45:11Z |
| format | Article |
| id | doaj.art-e0dcd06f577b4b4b9b9cf767f2cca35a |
| institution | Directory Open Access Journal |
| issn | 1177-8881 |
| language | English |
| last_indexed | 2024-12-10T14:45:11Z |
| publishDate | 2015-08-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | Drug Design, Development and Therapy |
| spelling | doaj.art-e0dcd06f577b4b4b9b9cf767f2cca35a2022-12-22T01:44:36ZengDove Medical PressDrug Design, Development and Therapy1177-88812015-08-012015default4883489623311High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumoniaYang XJWang YBZhou ZWWang GWWang XHLiu QFZhou SFWang ZHXiao-Jun Yang,1,* Yan-Bo Wang,2,3,* Zhi-Wei Zhou,4,* Guo-Wei Wang,2 Xiao-Hong Wang,1 Qing-Fu Liu,1 Shu-Feng Zhou,4 Zhen-Hai Wang2,3 1Department of Intensive Care Unit, 2Neurology Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People’s Republic of China; 3Key Laboratory of Brain Diseases of Ningxia, Yinchuan, Ningxia, People’s Republic of China; 4Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA *These authors contributed equally to this work Abstract: Ventilator-associated pneumonia (VAP) is a life-threatening disease that is associated with high rates of morbidity and likely mortality, placing a heavy burden on an individual and society. Currently available diagnostic and therapeutic approaches for VAP treatment are limited, and the prognosis of VAP is poor. The present study aimed to reveal and discriminate the identification of the full spectrum of the pathogens in patients with VAP using high-throughput sequencing approach and analyze the species richness and complexity via alpha and beta diversity analysis. The bronchoalveolar lavage fluid samples were collected from 27 patients with VAP in intensive care unit. The polymerase chain reaction products of the hypervariable regions of 16S rDNA gene in these 27 samples of VAP were sequenced using the 454 GS FLX system. A total of 103,856 pyrosequencing reads and 638 operational taxonomic units were obtained from these 27 samples. There were four dominant phyla, including Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. There were 90 different genera, of which 12 genera occurred in over ten different samples. The top five dominant genera were Streptococcus, Acinetobacter, Limnohabitans, Neisseria, and Corynebacterium, and the most widely distributed genera were Streptococcus, Limnohabitans, and Acinetobacter in these 27 samples. Of note, the mixed profile of causative pathogens was observed. Taken together, the results show that the high-throughput sequencing approach facilitates the characterization of the pathogens in bronchoalveolar lavage fluid samples and the determination of the profile for bacteria in the bronchoalveolar lavage fluid samples of the patients with VAP. This study can provide useful information of pathogens in VAP and assist clinicians to make rational and effective therapeutic decisions. Keywords: bioinformatics, drug resistance, OTU, PCR, DNA sequencinghttp://www.dovepress.com/1high-throughput-sequencing-of-16s-rdna-amplicons-characterizes-b-peer-reviewed-article-DDDT |
| spellingShingle | Yang XJ Wang YB Zhou ZW Wang GW Wang XH Liu QF Zhou SF Wang ZH High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia Drug Design, Development and Therapy |
| title | High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia |
| title_full | High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia |
| title_fullStr | High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia |
| title_full_unstemmed | High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia |
| title_short | High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator-associated pneumonia |
| title_sort | high throughput sequencing of 16s rdna amplicons characterizes bacterial composition in bronchoalveolar lavage fluid in patients with ventilator associated pneumonia |
| url | http://www.dovepress.com/1high-throughput-sequencing-of-16s-rdna-amplicons-characterizes-b-peer-reviewed-article-DDDT |
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