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Establishment of a Loop-mediated Isothermal Amplification Method for Rice Bacterial Leaf Brown Spot Disease
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作者 Zhang Jun-hua Wang Liang +8 位作者 Zhang Yao Ni Zhe Xu Xiao-feng Yang Ming-xiu Peng Li-li Yang Xin Wang Yi-han Jiang Xiao-jiao Haseeb Younis 《Journal of Northeast Agricultural University(English Edition)》 CAS 2023年第1期13-19,共7页
Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to es... Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice. 展开更多
关键词 Pseudomonas syringae pv.syringae loop-mediated isothermal amplification a rapid detection method
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Development and Evaluation of a Loop-mediated Isothermal Amplification(LAMP) Assay for Rapid Detection of Chinese Giant Salamander Ranavirus 被引量:3
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作者 Yi GENG Xingxing LIU +5 位作者 Yan ZHOU Kaiyu WANG Xi PENG Zhijun ZHONG Xiaoli HUANG Defang CHEN 《Asian Herpetological Research》 SCIE CSCD 2015年第1期59-65,共7页
Loop-mediated isothermal ampliifcation (LAMP) is a novel nucleic acid diagnostic method that can amplify rapidly a target template under isothermal conditions. In this study, a LAMP assay for rapid detection of Chin... Loop-mediated isothermal ampliifcation (LAMP) is a novel nucleic acid diagnostic method that can amplify rapidly a target template under isothermal conditions. In this study, a LAMP assay for rapid detection of Chinese giant salamander ranavirus(CGSRV) was developed from culture isolates and clinical samples. The LAMP assay was developed by designing one set of four speciifc primers, targeting the major capsid protein (MCP) gene of CGSRV. Reaction time and temperature were optimal for 40 min at 62°C. The developed LAMP assay is speciifc and highly sensitive for CGSRV detection, the detection limit could reach about 5 copies of cloned viral genomic fragments. The sensitivity of the LAMP assay was about 1000 and 10-fold higher than that of both conventional and nested PCR, respectively. The LAMP ampliifcation produces a typical ladder-like pattern of products on an agarose gel that can be visually inspected after addition of ethidium bromide. The LAMP assay was evaluated further with clinical samples, and the results indicated the suitability and simplicity of the test as a rapid diagnostic tool for the detection of CGSRV. 展开更多
关键词 CGSRV loop-mediated isothermal ampliifcation (lamp RANAVIRUS Chinese giant salamander
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One New Method of Nucleic Acid Amplification-Loop-mediated Isothermal Amplification of DNA 被引量:9
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作者 Xue-en FANG Jian LI Qin CHEN 《Virologica Sinica》 SCIE CAS CSCD 2008年第3期167-172,共6页
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method, which amplifies DNA with high specificity, sensitivity, rapidity and efficiency under isothermal conditions using a set of fo... Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method, which amplifies DNA with high specificity, sensitivity, rapidity and efficiency under isothermal conditions using a set of four specially designed primers and a Bst DNA polymerase with strand displacement activity. The basic principle, characteristics, development of LAMP and its applications are summarized in this article. 展开更多
关键词 Nucleic acid amplification loop-mediated isothermal amplification (lamp APPLICATION
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The Development of a Loop-Mediated Isothermal Amplification (LAMP) Procedure for Plague Diagnostic 被引量:1
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作者 Mariana de Lira Nunes Carina Lucena Mendes-Marques +1 位作者 Alzira Maria Paiva de Almeida Nilma Cintra Leal 《American Journal of Analytical Chemistry》 2014年第16期1069-1077,共9页
Plague caused by Yersinia pestis is one of the infectious diseases subject to the International Health Regulations (IHR). Permanent monitoring of the focal plague areas is mandatory in order to enable prompt control m... Plague caused by Yersinia pestis is one of the infectious diseases subject to the International Health Regulations (IHR). Permanent monitoring of the focal plague areas is mandatory in order to enable prompt control measures to prevent the spread of the disease. Therefore, the availability of efficient diagnosis tests is of paramount importance. Here, we describe a loop-mediated isothermal amplification (LAMP)-based procedure for rapid Y. pestis detection. We constructed a set of LAMP primers, which were used in assays to establish the reaction conditions that would lead to the quick visualization of the results by evaluating the test tube with the naked eye. The primers were specifically designed to target the caf1 gene located on pFra/Tox (pMT), a prototypical plasmid of Y. pestis. The LAMP procedure was performed at 65&deg;C for 45 min in a water bath and allowed for the detection of at least 10 pg of bacterial DNA. Due to its simplicity, specificity, sensitivity and rapidity, the LAMP technique is an additional tool that may be implemented in routine plague diagnoses, especially in emergencies. 展开更多
关键词 PLAGUE YERSINIA PESTIS Diagnosis Tests loop-mediated isothermal amplification (lamp)
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Development of a Loop-Mediated Isothermal Amplification Assay for Porcine Circovirus Type 2 被引量:3
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作者 Ye-bing Liu Lei Zhang +2 位作者 Qin-hong Xue Yi-bao Ning Zhi-gang Zhang 《Virologica Sinica》 SCIE CAS CSCD 2011年第3期214-220,共7页
In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBan... In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10-5 ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2. 展开更多
关键词 Porcine circovirus type 2 (PCV2) loop-mediated isothermal amplification (lamp Virus detection
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Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification 被引量:2
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作者 高宏伟 李富花 +2 位作者 张晓军 王兵 相建海 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第1期62-66,共5页
Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine... Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification (LAMP) assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase (empA) gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field. 展开更多
关键词 loop-mediated isothermal amplification (lamp detection assay empA gene Vibrio anguillarum
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Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification 被引量:1
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作者 ZHANG Fengying SHI Yanhong +2 位作者 JIANG Keji XU Zhaoli MA Lingbo 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2012年第2期139-146,共8页
A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (... A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum. 展开更多
关键词 Alexandrium eatenella Alexandrium minutum detection loop-mediated isothermal amplification (lamp ribosomal DNA internal transcribed spacer (ITS)
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Efficient detection of pathogen virus in sand dabs,Paralichthys olivaceus using loop-mediated isothermal amplification(LAMP) 被引量:1
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作者 HWANG Jinik PARK So Yun +3 位作者 SUH Sung-Suk PARK Mirye LEE Sukchan LEE Taek-Kyun 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2016年第8期44-50,共7页
Viral hemorrhagic septicemia virus(VHSV) and marine birnavirus(MABV) are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economi... Viral hemorrhagic septicemia virus(VHSV) and marine birnavirus(MABV) are the causative pathogens for some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogen infection. The loop-mediated isothermal amplification(LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools, as it is simple, quick, and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fish was developed. Using a set of synthesized primers matching a specific region of the genome, the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficient detection of VHSV and MABV infection in the wild fish, Paralichthys olivaceus. Our results demonstrate the simplicity and convenience of this method for the detection of viral infection in aquatic organisms. 展开更多
关键词 viral hemorrhagic septicaemia virus(VHSV) marine birnavirus(MABV) polymerase chain reaction loop-mediated isothermal amplification
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Preliminary investigation and detection based on loop-mediated isothermal amplification(LAMP)of phytoplasmas associated with diseases in B.napus L.
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作者 Yancheng Wen Shufen Zhang +8 位作者 Junping He Dongfang Cai Jiacheng Zhu Jianping Wang Jinhua Cao Kun Hu Lei Zhao Dongguo Wang Yizi Liu 《Oil Crop Science》 CSCD 2022年第4期219-224,共6页
In the last decade,some disease occurred on our experimental farms that had caused serious losses.They were not caused by fungi,bacteria or viruses.By loop-mediated isothermal amplification(LAMP)technique,the detectio... In the last decade,some disease occurred on our experimental farms that had caused serious losses.They were not caused by fungi,bacteria or viruses.By loop-mediated isothermal amplification(LAMP)technique,the detection results pointed to the possible pathogen as phytoplasma.The investigation results implied that phytoplasmas could cause more than 13 kinds of symptoms in almost all parts of plants in B.napus L.,including witches’broom,multi-stems,aggregate main inflorescences,and flat stems.The incidences of these phytoplasma-associated diseases in our experimental farms rose from 1.61%in 2010 to 6.00%in 2021.Some phytoplasma infected plants died without any growing points.These studies would be helpful for detecting phytoplasmas diseases,selecting disease resistant germplasm and improving varieties with disease resistances in B.napus L. 展开更多
关键词 B.napus L. Phytoplasma associated disease PATHOGEN DETECTION loop-mediated isothermal amplification (lamp)
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Smartphone-Imaged HIV-1 Reverse-Transcription Loop-Mediated Isothermal Ampliflcation(RT-LAMP) on a Chip from Whole Blood 被引量:9
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作者 Gregory L.Damhorst Carlos Duarte-Guevara +3 位作者 Weili Chen Tanmay Ghonge Brian T.Cunningham Rashid Bashir 《Engineering》 SCIE EI 2015年第3期324-335,共12页
Viral load measurements are an essential tool for the long-term clinical care of human immunodeficiency virus (HIV)-positive individuals. The gold standards in viral load instrumentation, however, are still too limi... Viral load measurements are an essential tool for the long-term clinical care of human immunodeficiency virus (HIV)-positive individuals. The gold standards in viral load instrumentation, however, are still too limited by their size, cost, and sophisticated operation for these measurements to be ubiquitous in remote settings with poor healthcare infrastructure, including parts of the world that are disproportionately affected by HIV infection. The challenge of developing a point-of-care platform capable of making viral load more accessible has been frequently approached but no solution has yet emerged that meets the practical requirements of low cost, portability, and ease-of-use. In this paper, we perform reverse-transcription loop-mediated isothermal amplification (RT-LAMP) on minimally processed HIV-spiked whole blood samples with a microfluidic and silicon microchip platform, and perform fluorescence measurements with a consumer smartphone. Our integrated assay shows amplification from as few as three viruses in a - 60 nL RT- LAMP droplet, corresponding to a whole blood concentration of 670 viruses per μL of whole blood. The technology contains greater power in a digital RT-LAMP approach that could be scaled up for the determination of viral load from a finger prick of blood in the clinical care of HIV-positive individuals. We demonstrate that all aspects of this viral load approach, from a drop of blood to imaging the RT-LAMP reaction, are compatible with lab-on-a-chip components and mobile instrumentation. 展开更多
关键词 human immunodeficiency virus (HIV) viral load loop-mediated isothermal amplification SMARTPHONE POINT-OF-CARE
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Rapid and Sensitive Detection of PRRSV by a Reverse Transcription-Loop-mediated Isothermal Amplification Assay 被引量:7
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作者 Lei Zhang Ye-bing Liu +2 位作者 Lei Chen Jian-huan Wang Yi-bao Ning 《Virologica Sinica》 SCIE CAS CSCD 2011年第4期252-259,共8页
A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive an... A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic,prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV) strains.As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR),the RT-LAMP method only used a turbidimeter,exhibited a detection limit corresponding to a 10-4 dilution of template RNA extracted from 250 μL of 105 of the 50% tissue culture infective dose (TCID50) of PRRSV-containing cells,and no cross-reactivity was observed with other related viruses including porcine circovirus type 2,swine influenza virus,porcine rotavirus and classical swine fever virus.From forty-two field samples,33 samples in the RT-LAMP assay was detected positive,whereas three of which were not detected by RT-PCR.Furthermore,in 33 strains of PRRSV,an identical detection rate was observed with the RT-LAMP assay to what were isolated using porcine alveolar macrophages.These findings demonstrated that the RT-LAMP assay has potential clinical applications for the detection of highly pathogenic PRRSV isolates,especially in developing countries. 展开更多
关键词 Reverse transcription loop-mediated isothermal amplification (RT-lamp) Porcine reproductive and respiratory syndrome virus (PRRSV) Clinical diagnosis Virus detection
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Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification assays for Rapid Detection of Human Enterovirus 71 and Coxsackievirus A16 in Clinical Samples 被引量:9
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作者 Hong Zhang Kai Nie +8 位作者 Yunzhi Liu Le Luo Wei Huang Shuaifeng Zhou Mengjie Yang Yu Chen Jianmin Luo Lidong Gao Xuejun Ma 《Advances in Infectious Diseases》 2012年第4期110-118,共9页
A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was perfor... A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16. The detection limits of RT-LAMP assays for both EV71 and CVA16 were 0.1 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain. The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19). In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (99.6%) of the specimens. Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16. In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions. 展开更多
关键词 Human ENTEROVIRUS 71 Coxsackievirus A16 REVERSE TRANSCRIPTION loop-mediated isothermal amplification
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Development and Evaluation of a Loop-mediated Isothermal Amplification Assay for the Rapid Detection and Identification of Pectobacterium carotovorum on Celery in the Field 被引量:3
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作者 Yanxia Shi Zhiwen Jin +4 位作者 Xianglong Meng Lixue Wang Xuewen Xie Ali Chai Baoju Li 《Horticultural Plant Journal》 SCIE 2020年第5期313-320,共8页
Pectobacterium carotovorum is the causal agent of bacterial soft rot in a wide range of vegetable host species.Once P.carotovorum infects the plant,the spread of the disease is difficult to control.In this study,a rap... Pectobacterium carotovorum is the causal agent of bacterial soft rot in a wide range of vegetable host species.Once P.carotovorum infects the plant,the spread of the disease is difficult to control.In this study,a rapid and sensitive method based on loop-mediated isothermal amplification(LAMP)was developed for detecting P.carotovorum in celery with soft rot using a primer set designed from the pmrA conserved sequence of P.carotovorum.The specificity of the LAMP primer set for P.carotovorum was extensively validated on both P.carotovorum strains and nontarget strains.The sensitivity was 1 pg of P.carotovorum genomic DNA,which demonstrated 10 times more sensitive than the conventional PCR assay.LAMP was also used to detect P.carotovorum in bacterial suspension.The lowest detection concentration was 104 CFU·mL^−1.In addition,a LAMP assay,in conjunction with a crude DNA extraction method,was successfully performed on P.carotovorum-infected samples derived from both artificially and naturally infected plants.In summary,the LAMP assay established in this study constitutes a simple,sensitive,and rapid method for the detection of P.carotovorum,and has potential application in the control of celery soft rot disease through early detection. 展开更多
关键词 Bacterial soft rot P.carotovorum loop-mediated isothermal amplification(lamp) CELERY pmrA gene field detection
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Development of loop-mediated isothermal amplification for rapid detection of Entamoeba histolytica 被引量:2
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作者 Windell L.Rivera Vanissa A.Ong 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2013年第6期457-461,共5页
Objective:To develop a loop-mediated isothermal amplification(LAMP) assay for the detection of Entamoeba histolytica(E.histolytica),the causative agent of amebiasis.Methods:The LAMP primer set was designed from E.hist... Objective:To develop a loop-mediated isothermal amplification(LAMP) assay for the detection of Entamoeba histolytica(E.histolytica),the causative agent of amebiasis.Methods:The LAMP primer set was designed from E.histolytica hemolysin gene HLY6.Genomic DNA of E.histolytica trophozoites strain HK9 was used to optimize the LAMP mixture and conditions.Amplification of DNA in the LAMP mixture was monitored through visual inspection for turbidity of the LAMP mix as well as addition of fluorescent dye.Results:Positive LAMP reactions turned turbid while negative ones remained clear.Upon addition of a fluorescent dye,all positive reactions turned green while the negative control remained orange under ambient light After elecrophoresis in 1.5% agarose gels,a ladder of multiple bands of different sizes can be oliserved in positive samples while no bands were detected in the negative control.The sensitivity of the assay was found to be S parasites per reaction which corresponds to approximately 1S.8 ng/μL DNA.The specificity of the assay was verified by the absence of amplified products when DNA from other gastrointestinal parasites such as the morphologically similar but non-pathogenic species,Entamoeba dispar. and other diarrhea-causing organisms such as Blastocystis hominis and Escherichia coli were used.Conclusions:The I.AMP assay we have developed enables the detection of E.histolytica with rapidity and ease,therefore rendering it is suitable for laboratory and field diagnosis of amebiasis. 展开更多
关键词 AMEBIASIS Diagnosis DNA ENTAMOEBA HISTOLYTICA HEMOLYSIN gene loop-mediated isothermal amplification assay
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Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification 被引量:2
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作者 Yuting Shang Shuzhen Cai +10 位作者 Qinghua Ye Qingping Wu Yanna Shao Xiaoying Qu Xinran Xiang Baoqing Zhou Yu Ding Moutong Chen Liang Xue Honghui Zhu Jumei Zhang 《Engineering》 SCIE EI CAS 2022年第12期62-70,共9页
Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined ... Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)detection.Quantum dot nanobeads(QBs)served as fluorescence reporters,providing good detection efficiency.The customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product capture.Detection was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)line.The linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test strips.ST specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and SD-LAMP.We detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis. 展开更多
关键词 Salmonella Typhimurium Quantum dot nanobeads Lateral flow immunoassay strip loop-mediated isothermal amplification Strand displacement probe
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Reverse Transcription-loop-mediated Isothermal Amplification for Detection of Maize Chlorotic Mottle Virus 被引量:4
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作者 Ying XU Yafei XU +2 位作者 Yongfeng LIU Zhijun QIU Wei ZHENG 《Agricultural Science & Technology》 CAS 2017年第1期123-126,共4页
Maize chlorotic mottle virus (MCMV) is a quarantine pest as approved by Chinese government: A rapid, sensitive and specific MCMV detection method using reverse transcription-loop-mediated isothermal amplification ... Maize chlorotic mottle virus (MCMV) is a quarantine pest as approved by Chinese government: A rapid, sensitive and specific MCMV detection method using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was established in this study. Based on the sequence of MCMV coat protein coding gene, 3 sets of primers were designed and specificity test showed that the second set of primers was specific to MCMV, Similar sensitivities were observed on RT-LAMP and RT-PCR, except that RT-LAMP was quicker, and the reaction could be finished within 1 h. In addition, the presence or absence of the fluorescence under daylight allows naked easy detection of the amplification of MCMV genomic RNA using calcein. The RT-LAMP assay was applied successfully to detect MCMV in maize seeds, and the result by the addition of calcein was consistent with the result detected by the real time turbidimeter. The method is rapid, specific, sensitive without the need for complicated equipment, and is suitable for rapid field detection of MCMV. 展开更多
关键词 Maize chlorotic mottle virus (MCMV) Reverse transcription loop-mediated isothermal amplification (RT-lamp) DETECTION
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Detection of Staphylococcus aureus in Dairy Food by Loop-mediated Isothermal Amplification 被引量:2
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作者 Cheng Xiao Wang Yongxin +4 位作者 An Hong Liu Juanjuan Zhang Bo Jia Zhen Cheng Jian 《Animal Husbandry and Feed Science》 CAS 2014年第3期127-130,共4页
[ Objective ] This study was to investigate the creditability of applying loop-mediated isothermal amplification method (LAMP) to detect Staphylococcus aureus in dairy products. [ Methods] The primers for heat resis... [ Objective ] This study was to investigate the creditability of applying loop-mediated isothermal amplification method (LAMP) to detect Staphylococcus aureus in dairy products. [ Methods] The primers for heat resistant nuclease gene (nuc) of Staphylococcus aureus were designed for establishing the LAMP method for rapidly detecting Staphylococcus aureus. [ Results ] The results of LAMP detection on Staphylococcus aureus in various dairy products were completely identical with that by bacterial isolation test; meanwhile it has a high specificity and a 10-fold sensitivity over the hemi-nested PCR. [ Conclusion] LAMP can be used for the detection of Staphylococcus aureus in dairy products. 展开更多
关键词 Staphylococcus aureus Nuc gene loop-mediated isothermal amplification
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Development of an in situ loop-mediated isothermal amplification technique for chromosomal localization of DNA sequences 被引量:1
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作者 孟庆磊 王师 +2 位作者 张玲玲 黄晓婷 包振民 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2013年第1期128-133,共6页
In situ loop-mediated isothermal amplification (in situ LAMP) combines in situ hybridization and loop-mediated isothermal amplification (LAMP) techniques for chromosomal localization of DNA sequences. In situ LAMP... In situ loop-mediated isothermal amplification (in situ LAMP) combines in situ hybridization and loop-mediated isothermal amplification (LAMP) techniques for chromosomal localization of DNA sequences. In situ LAMP is a method that is generally more specific and sensitive than conventional techniques such as fluorescence in situ hybridization (FISH), primed in situ labeling (PRINS), and cycling primed in situ labeling (C-PRINS). Here, we describe the development and application of in situ LAMP to identify the chromosomal localization of DNA sequences. To benchmark this technique, we successfully applied this technique to localize the major ribosomal RNA gene on the chromosomes of the Zhikong scallop ( Chlarnys farreri). 展开更多
关键词 chromosomal localization in situ loop-mediated isothermal amplification (in situ lamp) major rRNA Chlamys farreri
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Rapid,specific and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification targeted to vvhA gene 被引量:1
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作者 ZHANG Lina WANG Mingyi +5 位作者 CONG Dianxia DING Shuyan CONG Rinan YUE Jinyong GENG Jianli HU Chengjin 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2018年第4期83-88,共6页
Vibrio vulnificus is an estuarine bacterial pathogen for human.The rapid,specific and sensitive detection of V.vulnificus is urgently needed for early disease diagnosis and timely treatment of V.vulnificus infection.I... Vibrio vulnificus is an estuarine bacterial pathogen for human.The rapid,specific and sensitive detection of V.vulnificus is urgently needed for early disease diagnosis and timely treatment of V.vulnificus infection.In the study,a loop-mediated isothermal amplification(LAMP) technique was developed for V.vulnificus detection with a set of primers,composed of two out primers and two inner primers targeted to vvh A gene.The optimal amplification temperature was 63°C and the reaction only took 35 min.The amplification products could not only be detected by agarose gel electrophoresis with ladder-like pattern bands,but also could be visualized using calcein with naked eye directly.Forty-five strains were tested for the specificity of LAMP assay,and all the V.vulnificus strains were identified correctly while other strains were negative results.The sensitive of the new LAMP assay was 100-fold more sensitive than the conventional PCR.Meanwhile,all the V.vulnificus strains were detected correctly in spiked,clinical and environmental samples by the new LAMP assay.Compared with other well-known techniques,the new LAMP assay targeted to vvh A gene was extremely rapid,simple,sensitive and specific for V.vulnificus identification. 展开更多
关键词 Vibrio uulnificus loop-mediated isothermal amplification vvhA gene
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Combination of Loop-Mediated Isothermal Amplification Assay and Nested PCR for Detection of Borrelia burgdorferi sensu lato in Human Serum Samples 被引量:1
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作者 ZHANG Liu Li HOU Xue Xia +3 位作者 GENG Zhen LOU Yong Liang WAN Kang Lin HAO Qin 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2015年第4期312-315,共4页
A set of universal loop-mediated isothermal amplification (LAMP) primers targeting the flo gene was designed to detect Borrelia burgdorferi sensu lato (B. burgdorferi s.I.) in human samples. The sensitivity of LAM... A set of universal loop-mediated isothermal amplification (LAMP) primers targeting the flo gene was designed to detect Borrelia burgdorferi sensu lato (B. burgdorferi s.I.) in human samples. The sensitivity of LAMP was 20 copies/reaction, and the assay did not detect false positives among 11 other related bacteria. A positive LAMP result was obtained for 9 of the 24 confirmed cases and for 12 of 94 suspected cases. The positive rate of LAMP was the same as that of nested PCR. The LAMP is a useful diagnostic method that can be developed for rapid detection of B. burgdorferi s.I. in human sera. Combination of the LAMP and nested PCR was more sensitive for detecting B. burgdorferi s.I. in human serum samples. 展开更多
关键词 PCR lamp Combination of loop-mediated isothermal amplification Assay and Nested PCR for Detection of Borrelia burgdorferi sensu lato in Human Serum Samples
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