ddPCR surpasses classical qPCR technology in quantitating bacteria and fungi in the environment
文献类型：期刊论文
| 作者 | Wang, Danrui ; Wang, Shang; Du, Xiongfeng; He, Qing; Liu, Yue; Wang, Zhujun; Feng, Kai; Li, Yan; Deng, Ye
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| 刊名 | MOLECULAR ECOLOGY RESOURCES
 |
| 出版日期 | 2022 |
| 卷号 | 22期号:7页码:2587-2598 |
| ISSN号 | 1755-098X |
| 关键词 | DROPLET DIGITAL PCR MICROBIAL COMMUNITIES INHIBITION QUANTIFICATION DNA BIOMASS CARBON DILUTION IMPACTS TAQMAN |
| 英文摘要 | Quantitative real-time PCR (qPCR) has been widely used in quantifying bacterial and fungal populations in various ecosystems, as well as the fungi to bacteria ratio (F:B ratio). Recently, researchers have begun to apply droplet digital PCR (ddPCR) to this area; however, no study has systematically compared qPCR and ddPCR for quantitating both bacteria and fungi in environmental samples at the same time. Here, we designed probe-primer pair combinations targeting the 16S rRNA gene and internal transcribed spacer (ITS) for the detection of bacteria and fungi, respectively, and tested both SYBR Green and TaqMan approaches in qPCR and ddPCR methods for mock communities and in real environmental samples. In mock communities, the quantification results of ddPCR were significantly closer to expected values (p < .05), and had smaller coefficients of variations (p < .05) than qPCR, suggesting ddPCR was more accurate and repeatable. In environmental samples, ddPCR consistently quantified ITS and 16S rRNA gene concentrations in all four habitats without abnormal overestimation or underestimation, and the F:B ratio obtained by ddPCR was consistent with phospholipid fatty acid analysis. Our results indicated that ddPCR had better precision, repeatability, sensitivity, and stability in bacterial and fungal quantitation than qPCR. Although ddPCR has high cost, complicated processes and restricted detection range, it shows insensitivity to PCR inhibitors and the potential of quantifying long target fragments. We expect that ddPCR, which is complementary to qPCR, will contribute to microbial quantification in environmental monitoring and evaluation. |
| 源URL | [https://ir.rcees.ac.cn/handle/311016/47776]  |
| 专题 | 生态环境研究中心_中国科学院环境生物技术重点实验室 |
| 通讯作者 | Deng, Ye |
| 作者单位 | 1.Sichuan Univ, West China Hosp Stomatol, Natl Clin Res Ctr Oral Dis, State Key Lab Oral Dis, Chengdu, Peoples R China 2.Univ Chinese Acad Sci, Coll Resources & Environm, Beijing, Peoples R China 3.Chinese Acad Sci, Res Ctr Ecoenvironm Sci, CAS Key Lab Environm Biotechnol, Beijing 100085, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Danrui,Wang, Shang,Du, Xiongfeng,et al. ddPCR surpasses classical qPCR technology in quantitating bacteria and fungi in the environment[J]. MOLECULAR ECOLOGY RESOURCES,2022,22(7):2587-2598. |
| APA | Wang, Danrui.,Wang, Shang.,Du, Xiongfeng.,He, Qing.,Liu, Yue.,...&Deng, Ye.(2022).ddPCR surpasses classical qPCR technology in quantitating bacteria and fungi in the environment.MOLECULAR ECOLOGY RESOURCES,22(7),2587-2598. |
| MLA | Wang, Danrui,et al."ddPCR surpasses classical qPCR technology in quantitating bacteria and fungi in the environment".MOLECULAR ECOLOGY RESOURCES 22.7(2022):2587-2598. |
入库方式： OAI收割
来源：生态环境研究中心
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