Helix loop-mediated isothermal amplification of nucleic acids
Mao, Rui1,2; Qi, Lifei1,2; Wang, Zhuo1; Liu, Hongtao1,3; Du, Yuguang1
Source PublicationRSC ADVANCES
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.
Funding ProjectChinese Academy of Sciences[CXJJ-15Z012]
WOS Research AreaChemistry
WOS SubjectChemistry, Multidisciplinary
Funding OrganizationChinese Academy of Sciences
WOS IDWOS:000433391600038
Citation statistics
Document Type期刊论文
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.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Mao, Rui]'s Articles
[Qi, Lifei]'s Articles
[Wang, Zhuo]'s Articles
Baidu academic
Similar articles in Baidu academic
[Mao, Rui]'s Articles
[Qi, Lifei]'s Articles
[Wang, Zhuo]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Mao, Rui]'s Articles
[Qi, Lifei]'s Articles
[Wang, Zhuo]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.