CAS OpenIR
Helix loop-mediated isothermal amplification of nucleic acids
Mao, Rui1,2; Qi, Lifei1,2; Wang, Zhuo1; Liu, Hongtao1,3; Du, Yuguang1
2018
Source PublicationRSC ADVANCES
ISSN2046-2069
Volume8Issue:34Pages:19098-19102
AbstractIsothermal nucleic acid amplification has played a key role in the point of care test (POCT). In this study, a helix loop-mediated isothermal amplification (HAMP) method with high specificity, efficiency and rapidity was developed. The MERS-Cov orf1b gene was chosen for the validation and optimization of HAMP. The HAMP analysis was performed at a constant temperature of 61-65 degrees C and yielded a self-primed spiral structure with no introduction of exogenous gene sequence by two pairs of specially designed primers. The primers for helix loop formation were composed of two complementary primers including the helix forward primer and the helix reverse primer, the 3 ends of which were complementary to their respective target nucleic acids. HAMP assay can be monitored by fluorescence signals with the addition of Eva Green in the reaction mixture. In addition, an accelerated HAMP was developed after the addition of acceleration probe, which could be finished within 75 min with a sensitivity of 10 copies per reaction. Further, a reverse transcription-HAMP (RT-HAMP) was proven to be feasible for RNA detection by combining the reverse transcriptase with DNA polymerase. Finally, both the HAMP and RT-HAMP assay were visually conducted by using Hydroxynaphthol blue (HNB) as a chromogenic indicator. All in all, it is suggested that the HAMP assay would have great potential in POCT applications.
DOI10.1039/c8ra01201f
Language英语
WOS KeywordINFECTIOUS-DISEASES ; DNA ; DIAGNOSTICS ; LAMP
Funding ProjectChinese Academy of Sciences[CXJJ-15Z012]
WOS Research AreaChemistry
WOS SubjectChemistry, Multidisciplinary
Funding OrganizationChinese Academy of Sciences
WOS IDWOS:000433391600038
PublisherROYAL SOC CHEMISTRY
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/24794
Collection中国科学院过程工程研究所
Corresponding AuthorWang, Zhuo; Liu, Hongtao; Du, Yuguang
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Key Lab Biopharmaceut Prod & Formulat Engn, State Key Lab Biochem Engn,PLA, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Zhengzhou Inst Emerging Ind Technol, Zhengzhou 450000, Henan, Peoples R China
Recommended Citation
GB/T 7714
Mao, Rui,Qi, Lifei,Wang, Zhuo,et al. Helix loop-mediated isothermal amplification of nucleic acids[J]. RSC ADVANCES,2018,8(34):19098-19102.
APA Mao, Rui,Qi, Lifei,Wang, Zhuo,Liu, Hongtao,&Du, Yuguang.(2018).Helix loop-mediated isothermal amplification of nucleic acids.RSC ADVANCES,8(34),19098-19102.
MLA Mao, Rui,et al."Helix loop-mediated isothermal amplification of nucleic acids".RSC ADVANCES 8.34(2018):19098-19102.
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