Development of a single-tube nested PCR-lateral flow biosensor assay for rapid and accurate detection of Alternaria panax Whetz

Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the eff...

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Veröffentlicht in:	PloS one Jg. 13; H. 11; S. e0206462
Hauptverfasser:	Wei, Shuqin, Sun, Yajuan, Xi, Guangsheng, Zhang, Huijuan, Xiao, Mingya, Yin, Rui
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	United States Public Library of Science 08.11.2018 Public Library of Science (PLoS)
Schlagworte:	Alternaria - genetics Alternaria - isolation & purification Assaying Biology and Life Sciences Biosensing Techniques - methods Biosensors Cultivation Deoxyribonucleic acid Diagnostic systems DNA DNA, Fungal - genetics Ecology and Environmental Sciences Economic impact Engineering and Technology Ginseng Hybridization Identification Limit of Detection Medicine and Health Sciences Methods Migration Nucleic Acid Hybridization Pathogens Pharmaceuticals Plant diseases Plant pathology Polymerase chain reaction Polymerase Chain Reaction - methods Research and Analysis Methods RNA, Ribosomal, 18S - genetics Time Factors China
ISSN:	1932-6203, 1932-6203
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Abstract	Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the effective prevention and control of further infection spread. In this work, a rapid and accurate molecular diagnostic method, a single-tube nested PCR-lateral flow biosensor assay (STNPCR-LFBA), was developed for rapid identification of Alternaria panax Whetz. The STNPCR-LFBA was 100 times more sensitive than the traditional PCR-LFBA. Besides that, the PCR product was checked by a lateral flow biosensor assay, which provided a basis for the migration of the detection technology to a point-of-care test (POCT) format. STNPCR-LFBA was specific to Alternaria panax Whetz, and no cross-reactions were observed in other non-target samples; the limit of detection was up to 0.01 pg of Alternaria panax Whetz genomic DNA. STNPCR-LFBA could also be used for specific identification of Alternaria panax Whetz in real samples. STNPCR-LFBA is useful for identifying Alternaria panax Whetz due to its rapidity, accuracy, and simple manipulation.
AbstractList	Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the effective prevention and control of further infection spread. In this work, a rapid and accurate molecular diagnostic method, a single-tube nested PCR-lateral flow biosensor assay (STNPCR-LFBA), was developed for rapid identification of Alternaria panax Whetz. The STNPCR-LFBA was 100 times more sensitive than the traditional PCR-LFBA. Besides that, the PCR product was checked by a lateral flow biosensor assay, which provided a basis for the migration of the detection technology to a point-of-care test (POCT) format. STNPCR-LFBA was specific to Alternaria panax Whetz, and no cross-reactions were observed in other non-target samples; the limit of detection was up to 0.01 pg of Alternaria panax Whetz genomic DNA. STNPCR-LFBA could also be used for specific identification of Alternaria panax Whetz in real samples. STNPCR-LFBA is useful for identifying Alternaria panax Whetz due to its rapidity, accuracy, and simple manipulation. Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the effective prevention and control of further infection spread. In this work, a rapid and accurate molecular diagnostic method, a single-tube nested PCR-lateral flow biosensor assay (STNPCR-LFBA), was developed for rapid identification of Alternaria panax Whetz. The STNPCR-LFBA was 100 times more sensitive than the traditional PCR-LFBA. Besides that, the PCR product was checked by a lateral flow biosensor assay, which provided a basis for the migration of the detection technology to a point-of-care test (POCT) format. STNPCR-LFBA was specific to Alternaria panax Whetz, and no cross-reactions were observed in other non-target samples; the limit of detection was up to 0.01 pg of Alternaria panax Whetz genomic DNA. STNPCR-LFBA could also be used for specific identification of Alternaria panax Whetz in real samples. STNPCR-LFBA is useful for identifying Alternaria panax Whetz due to its rapidity, accuracy, and simple manipulation.Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the effective prevention and control of further infection spread. In this work, a rapid and accurate molecular diagnostic method, a single-tube nested PCR-lateral flow biosensor assay (STNPCR-LFBA), was developed for rapid identification of Alternaria panax Whetz. The STNPCR-LFBA was 100 times more sensitive than the traditional PCR-LFBA. Besides that, the PCR product was checked by a lateral flow biosensor assay, which provided a basis for the migration of the detection technology to a point-of-care test (POCT) format. STNPCR-LFBA was specific to Alternaria panax Whetz, and no cross-reactions were observed in other non-target samples; the limit of detection was up to 0.01 pg of Alternaria panax Whetz genomic DNA. STNPCR-LFBA could also be used for specific identification of Alternaria panax Whetz in real samples. STNPCR-LFBA is useful for identifying Alternaria panax Whetz due to its rapidity, accuracy, and simple manipulation.
Author	Sun, Yajuan Xiao, Mingya Zhang, Huijuan Yin, Rui Wei, Shuqin Xi, Guangsheng
AuthorAffiliation	1 College of Agronomy, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China University of Helsinki, FINLAND 2 Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China 3 College of Traditional Chinese Medicine, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China 4 College of Biological and Pharmaceutical Engineering, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China
AuthorAffiliation_xml	– name: University of Helsinki, FINLAND – name: 2 Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China – name: 4 College of Biological and Pharmaceutical Engineering, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China – name: 1 College of Agronomy, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China – name: 3 College of Traditional Chinese Medicine, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China
Author_xml	– sequence: 1 givenname: Shuqin surname: Wei fullname: Wei, Shuqin – sequence: 2 givenname: Yajuan surname: Sun fullname: Sun, Yajuan – sequence: 3 givenname: Guangsheng surname: Xi fullname: Xi, Guangsheng – sequence: 4 givenname: Huijuan surname: Zhang fullname: Zhang, Huijuan – sequence: 5 givenname: Mingya surname: Xiao fullname: Xiao, Mingya – sequence: 6 givenname: Rui orcidid: 0000-0002-4852-5063 surname: Yin fullname: Yin, Rui
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Copyright	2018 Wei et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. 2018 Wei et al 2018 Wei et al
Copyright_xml	– notice: 2018 Wei et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. – notice: 2018 Wei et al 2018 Wei et al
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Snippet	Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic...
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SubjectTerms	Alternaria - genetics Alternaria - isolation & purification Assaying Biology and Life Sciences Biosensing Techniques - methods Biosensors Cultivation Deoxyribonucleic acid Diagnostic systems DNA DNA, Fungal - genetics Ecology and Environmental Sciences Economic impact Engineering and Technology Ginseng Hybridization Identification Limit of Detection Medicine and Health Sciences Methods Migration Nucleic Acid Hybridization Pathogens Pharmaceuticals Plant diseases Plant pathology Polymerase chain reaction Polymerase Chain Reaction - methods Research and Analysis Methods RNA, Ribosomal, 18S - genetics Time Factors
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Title	Development of a single-tube nested PCR-lateral flow biosensor assay for rapid and accurate detection of Alternaria panax Whetz
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