A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection

ABSTRACT Human adenoviruses (HAdVs) are common viruses that can cause local outbreaks in schools, communities and military camps, posing a huge threat to public health. An ideal POCT device for adenovirus detection in resource-limited settings is critical to control the spread of the virus. In this...

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Main Authors: Kui Sun, Xiaodong Yang, Yanan Wang, Qun Guan, Wenliang Fu, Chao Zhang, Qin Liu, Wenzheng An, Yongqi Zhao, Weiwei Xing, Donggang Xu
Format: Article
Language:English
Published: American Society for Microbiology 2023-06-01
Series:Microbiology Spectrum
Subjects:
Online Access:https://journals.asm.org/doi/10.1128/spectrum.05170-22
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author Kui Sun
Xiaodong Yang
Yanan Wang
Qun Guan
Wenliang Fu
Chao Zhang
Qin Liu
Wenzheng An
Yongqi Zhao
Weiwei Xing
Donggang Xu
author_facet Kui Sun
Xiaodong Yang
Yanan Wang
Qun Guan
Wenliang Fu
Chao Zhang
Qin Liu
Wenzheng An
Yongqi Zhao
Weiwei Xing
Donggang Xu
author_sort Kui Sun
collection DOAJ
description ABSTRACT Human adenoviruses (HAdVs) are common viruses that can cause local outbreaks in schools, communities and military camps, posing a huge threat to public health. An ideal POCT device for adenovirus detection in resource-limited settings is critical to control the spread of the virus. In this study, we developed an integrated and electricity-independent sample-to-answer system that can complete nucleic acid extraction, amplification, and detection at room temperature. This system is suitable for field and on-site detection because of its rapidity, sensitivity, lack of contamination, and lack of requirements of high-precision instruments and skilled technicians. It consists of two separate modules, ALP FINA (alkaline lysis with the paper-based filtration isolation of nucleic acid) and SV RPA (sealed and visual recombinase polymerase amplification). The extraction efficiency of ALP FINA can reach 48 to 84%, which is close to that of the conventional centrifuge column. The detection sensitivity of SV RPA is close to 10 copies/μL of AdvB and AdvE without aerosol contamination after repeated operations. When SV RPA was applied to the detection of nasopharyngeal swab samples of 19 patients who were infected with AdvB or AdvE as well as 10 healthy volunteers, its sensitivity and specificity reached 100%, respectively. IMPORTANCE HAdV infections are readily transmittable and, in some instances, highly contagious. Early and rapid diagnosis is essential for disease control. In this work, we developed a portable, disposable, and modularized sample-to-answer detection system for AdvB and AdvE, which rendered the entire test to be completely independent of electricity and other laboratory infrastructure. Thus, this detection system can be applied in resource-limited settings, and it has the potential to be further developed as an early diagnosis method in the field.
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spelling doaj.art-05c15e36d1aa489e8ad126cfb3b2fe282023-06-15T13:18:31ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-06-0111310.1128/spectrum.05170-22A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus DetectionKui Sun0Xiaodong Yang1Yanan Wang2Qun Guan3Wenliang Fu4Chao Zhang5Qin Liu6Wenzheng An7Yongqi Zhao8Weiwei Xing9Donggang Xu10Beijing Institute of Basic Medical Sciences, Beijing, ChinaDepartment of General Surgery, the First Medical Center of Chinese PLA General Hospital, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaThe Fifth Medical Center of Chinese PLA General Hospital, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaBeijing Institute of Basic Medical Sciences, Beijing, ChinaABSTRACT Human adenoviruses (HAdVs) are common viruses that can cause local outbreaks in schools, communities and military camps, posing a huge threat to public health. An ideal POCT device for adenovirus detection in resource-limited settings is critical to control the spread of the virus. In this study, we developed an integrated and electricity-independent sample-to-answer system that can complete nucleic acid extraction, amplification, and detection at room temperature. This system is suitable for field and on-site detection because of its rapidity, sensitivity, lack of contamination, and lack of requirements of high-precision instruments and skilled technicians. It consists of two separate modules, ALP FINA (alkaline lysis with the paper-based filtration isolation of nucleic acid) and SV RPA (sealed and visual recombinase polymerase amplification). The extraction efficiency of ALP FINA can reach 48 to 84%, which is close to that of the conventional centrifuge column. The detection sensitivity of SV RPA is close to 10 copies/μL of AdvB and AdvE without aerosol contamination after repeated operations. When SV RPA was applied to the detection of nasopharyngeal swab samples of 19 patients who were infected with AdvB or AdvE as well as 10 healthy volunteers, its sensitivity and specificity reached 100%, respectively. IMPORTANCE HAdV infections are readily transmittable and, in some instances, highly contagious. Early and rapid diagnosis is essential for disease control. In this work, we developed a portable, disposable, and modularized sample-to-answer detection system for AdvB and AdvE, which rendered the entire test to be completely independent of electricity and other laboratory infrastructure. Thus, this detection system can be applied in resource-limited settings, and it has the potential to be further developed as an early diagnosis method in the field.https://journals.asm.org/doi/10.1128/spectrum.05170-22RPAsample to answerhuman adenovirusesaerosol contaminationNAAT
spellingShingle Kui Sun
Xiaodong Yang
Yanan Wang
Qun Guan
Wenliang Fu
Chao Zhang
Qin Liu
Wenzheng An
Yongqi Zhao
Weiwei Xing
Donggang Xu
A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
Microbiology Spectrum
RPA
sample to answer
human adenoviruses
aerosol contamination
NAAT
title A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
title_full A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
title_fullStr A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
title_full_unstemmed A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
title_short A Novel Sample-to-Answer Visual Nucleic Acid Detection System for Adenovirus Detection
title_sort novel sample to answer visual nucleic acid detection system for adenovirus detection
topic RPA
sample to answer
human adenoviruses
aerosol contamination
NAAT
url https://journals.asm.org/doi/10.1128/spectrum.05170-22
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