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

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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.

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          Most cited references25

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          Medicinal plants and phytomedicines. Linking plant biochemistry and physiology to human health.

          D Briskin (2000)
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            Comparison of the pharmacological effects of Panax ginseng and Panax quinquefolium.

            Medical application of Panax ginseng was first found in "Shen-Nong Herbal Classic"around 200 AD Panax quinquefolium was first introduced in "Essential of Materia Medica" in 1694 in China. The most important bioactive components contained in P ginseng and P quinquefolium are ginseng saponins (GS). The contents of ginsenoside Rb1, Re, and Rd in P quinquefolium are higher than they are in P ginseng. In P ginseng, the contents of Rg1,Rb2, and Rc are higher than they are in P quinquefolium. P ginseng had a higher ratio of Rg1: Rb1, and which was lower in P quinquefolium. After steaming for several hours, the total GS will decrease. However, some ginsenosides (Rg2, 20R-Rg2, Rg3, Rh1 and Rh2) increase, while others (Rb1, Rb2, Rb3, Rc, Rd, Re, and Rg1) decrease. However, variation, especially in P quinquefolium, is high. P ginseng and P quinquefolium are general tonics and adaptogens. Rg1 and Rb1 enhance central nervous system (CNS) activities, but the effect of the latter is weaker. Thus, for the higher contents of Rg1, P ginseng is a stimulant, whereas the Rb1 contents of P quinquefolium are mainly calming to the CNS. Re, Rg1, panaxan A and B from P ginseng are good for diabetes. Re and Rg1 enhance angiogenesis, whereas Rb1, Rg3 and Rh2 inhibit it. Rh2, an antitumor agent, can be obtained from Rb1 by steaming. The content of Re in P quinquefolium are higher than in P ginseng by 3-4 times. The vasorelax, antioxidant, antihyperlipidemic, and angiogenic effects of Re are reported. Thus, for the CNS "hot," wound healing and hypoglycemic effects, P ginseng is better than P quinquefolium. For anticancer effects, P quinquefolium is better.
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              Trends in Ginseng Research in 2010

              A total of 470 papers directly related to research on the Panax species were retrieved by performing internet searches with the keywords Panax and ginseng as the search terms. The publications were categorized as follows: 399 research articles, 30 reviews, 30 meeting abstracts, 7 proceedings, and 4 letters. The majority of these publications were published by scientists from Korea (35.7%), China (32.3%), and the USA (11.3%). Scientists from a total of 29 nations were actively involved in conducting ginseng research. A total of 43.6% of the publications were categorized as pharmacodynamic studies. The effects of ginseng on cerebrovascular function and cancer were the two most common topics considered in the pharmacodynamic studies. More than half of the ginseng studies assessed the use of P. ginseng. A total of 23 countries participated in studies specifically related to P. ginseng, and more than 80% of these studies originated from Korea and China. A total of 50 topics within the pharmacodynamics category were examined in association with the use of P. ginseng.
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                Author and article information

                Contributors
                Role: Funding acquisitionRole: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: Methodology
                Role: Funding acquisition
                Role: MethodologyRole: Software
                Role: Data curationRole: Formal analysis
                Role: MethodologyRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                8 November 2018
                2018
                : 13
                : 11
                : e0206462
                Affiliations
                [1 ] College of Agronomy, Jilin College of Agricultural Science and Technology, Jilin, Jilin, China
                [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
                University of Helsinki, FINLAND
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-4852-5063
                Article
                PONE-D-18-22074
                10.1371/journal.pone.0206462
                6224276
                30408825
                bea1ac5b-5a25-4e33-95b4-ded6e057de38
                © 2018 Wei et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 25 July 2018
                : 13 October 2018
                Page count
                Figures: 6, Tables: 1, Pages: 11
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100003758, China Natural Science Foundation;
                Award ID: 31470420
                Award Recipient :
                This work was financially supported by the China Natural Science Foundation (31470420 to SW), http://www.nsfc.gov.cn/; Jilin Ginseng High Scientific and Technological Innovation Platform, Jilin Provincial Key Laboratory of Technological Innovation in Production and Utilization of Genuine Regional Drug, the Project of Scientific and Technological Innovation Team of Key Cultivation Technology of Changbai Mountain's genuine medicinal herbs, http://www.nsfc.gov.cn/.
                Categories
                Research Article
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Research and Analysis Methods
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Engineering and Technology
                Equipment
                Detectors
                Biosensors
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Fungal Pathogens
                Alternaria
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Fungal Pathogens
                Alternaria
                Biology and Life Sciences
                Mycology
                Fungal Pathogens
                Alternaria
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Molecular Probe Techniques
                Probe Hybridization
                Research and Analysis Methods
                Molecular Biology Techniques
                Molecular Probe Techniques
                Probe Hybridization
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Nested Polymerase Chain Reaction
                Research and Analysis Methods
                Molecular Biology Techniques
                Artificial Gene Amplification and Extension
                Polymerase Chain Reaction
                Nested Polymerase Chain Reaction
                Biology and Life Sciences
                Agriculture
                Agricultural Soil Science
                Ecology and Environmental Sciences
                Soil Science
                Agricultural Soil Science
                Biology and Life Sciences
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                All relevant data are within the paper and its Supporting Information files.

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