Diagnosis of maize lethal necrosis (MLN)-causing viruses is key in MLN surveillance programs and in testing seed for zero tolerance of Maize chlorotic mottle virus (MCMV) in seed lots. This is crucial for MLN management in farmers’ fields and in commercial seed fields. A customized MCMV detection assay that is specific, sensitive, affordable, and portable is therefore important for this task. Reverse Transcriptase – Recombinase Polymerase Amplification (RT-RPA) meets those conditions earlier described. RPA is a rapid isothermal nucleic acid amplification and detection platform that is based on patented Recombinase Polymerase Amplification (RPA) technology. In this study, a real time endpoint analysis and field deployable RT-RPA diagnostic method for the detection of MCMV was developed. RPA primer sets with their complementary probes were designed, synthesized and tested through a series of primer set evaluations to determine the most efficient primer sets. The primer sets targeted the MCMV genome at position 2765-2948 bp (MCMV_gp2 replicase gene). The parameters evaluated were sensitivity, specificity and reproducibility for the assay with remarkable results. The assay discriminated against other maize infecting viruses hence specific to MCMV. The assay takes only 20 min and its detection limit of 10 -4 is well comparable to RT-PCR and other molecular based detection assays. MCMV was also detected directly from leaf saps without the nucleic acid extraction step hence suitable for on-farm testing. RPA is a relatively inexpensive technique that requires minimal instrumentation. This assay is therefore suitable for the detection of MCMV in field surveys, routine MCMV testing for phytosanitary measures and in the seed certification procedures.
{"title":"Field deployable Reverse TranscriptaseRecombinase Polymerase Amplification (RT-RPA) for detection of Maize chlorotic mottle virus (MCMV)","authors":"M. F., G. M., M. S., M. X., N. B., M. S. L.","doi":"10.5897/jgmv2022.0082","DOIUrl":"https://doi.org/10.5897/jgmv2022.0082","url":null,"abstract":"Diagnosis of maize lethal necrosis (MLN)-causing viruses is key in MLN surveillance programs and in testing seed for zero tolerance of Maize chlorotic mottle virus (MCMV) in seed lots. This is crucial for MLN management in farmers’ fields and in commercial seed fields. A customized MCMV detection assay that is specific, sensitive, affordable, and portable is therefore important for this task. Reverse Transcriptase – Recombinase Polymerase Amplification (RT-RPA) meets those conditions earlier described. RPA is a rapid isothermal nucleic acid amplification and detection platform that is based on patented Recombinase Polymerase Amplification (RPA) technology. In this study, a real time endpoint analysis and field deployable RT-RPA diagnostic method for the detection of MCMV was developed. RPA primer sets with their complementary probes were designed, synthesized and tested through a series of primer set evaluations to determine the most efficient primer sets. The primer sets targeted the MCMV genome at position 2765-2948 bp (MCMV_gp2 replicase gene). The parameters evaluated were sensitivity, specificity and reproducibility for the assay with remarkable results. The assay discriminated against other maize infecting viruses hence specific to MCMV. The assay takes only 20 min and its detection limit of 10 -4 is well comparable to RT-PCR and other molecular based detection assays. MCMV was also detected directly from leaf saps without the nucleic acid extraction step hence suitable for on-farm testing. RPA is a relatively inexpensive technique that requires minimal instrumentation. This assay is therefore suitable for the detection of MCMV in field surveys, routine MCMV testing for phytosanitary measures and in the seed certification procedures.","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44002684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-01-01Epub Date: 2019-06-30DOI: 10.5897/JGMV2019.0073
L A O Awata, B E Ifie, P Tongoona, E Danquah, M B Jumbo, M Gowda, P W Marchelo-D'ragga, Chelang'at Sitonik, L M Suresh
Maize lethal necrosis (MLN) disease is new to Africa. First report was in Kenya in 2012, since then the disease has rapidly spread to most parts of eastern and central Africa region including Tanzania, Burundi, DRC Congo, Rwanda, Uganda, Ethiopia and similar symptoms were observed in South Sudan. Elsewhere, the disease was caused by infection of Maize Chlorotic Mottle Virus (MCMV) in combination with any of the potyviruses namely; maize dwarf mosaic virus (MDMV), sugarcane mosaic virus (SCMV) and tritimovirus wheat streak mosaic virus (WSMV). In Africa, the disease occurs due to combined infections of maize by MCMV and SCMV, leading to severe yield losses. Efforts to address the disease spread have been ongoing. Serological techniques including enzyme-linked immuno-sorbent assay (ELISA), polymerase chain reaction (PCR), genome-wide association (GWAS) mapping and next generation sequencing have been effectively used to detect and characterize MLN causative pathogens. Various management strategies have been adapted to control MLN including use of resistant varieties, phytosanitary measures and better cultural practices. This review looks at the current knowledge on MLN causative viruses, genetic architecture and molecular basis underlying their synergistic interactions. Lastly, some research gaps towards MLN management will be identified. The information gathered may be useful for developing strategies towards future MLN management and maize improvement in Africa.
玉米致死坏死病（MLN）是非洲的新病害。首次报告发生在 2012 年的肯尼亚，此后该病迅速蔓延到非洲东部和中部的大部分地区，包括坦桑尼亚、布隆迪、刚果（金）、卢旺达、乌干达和埃塞俄比亚，南苏丹也出现了类似症状。在其他地区，该病是由玉米萎黄斑驳病毒（MCMV）与玉米矮化花叶病毒（MDMV）、甘蔗花叶病毒（SCMV）和小麦条纹花叶病毒（WSMV）中的任何一种病毒感染引起的。在非洲，这种疾病是由玉米矮化花叶病毒（MCMV）和甘蔗花叶病毒（SCMV）共同感染玉米引起的，导致严重的产量损失。目前正在努力解决该疾病的传播问题。血清学技术，包括酶联免疫吸附试验（ELISA）、聚合酶链反应（PCR）、全基因组关联图谱（GWAS）和新一代测序，已被有效用于检测和鉴定 MLN 致病病原体。为控制 MLN，人们采用了各种管理策略，包括使用抗病品种、植物检疫措施和更好的栽培方法。本综述探讨了有关 MLN 致病病毒、遗传结构及其协同作用的分子基础的现有知识。最后，还将指出在多发性唇疱疹管理方面存在的一些研究空白。所收集的信息可能有助于为非洲未来的多瘤病毒管理和玉米改良制定战略。
{"title":"Maize lethal necrosis and the molecular basis of variability in concentrations of the causal viruses in co-infected maize plant.","authors":"L A O Awata, B E Ifie, P Tongoona, E Danquah, M B Jumbo, M Gowda, P W Marchelo-D'ragga, Chelang'at Sitonik, L M Suresh","doi":"10.5897/JGMV2019.0073","DOIUrl":"10.5897/JGMV2019.0073","url":null,"abstract":"<p><p>Maize lethal necrosis (MLN) disease is new to Africa. First report was in Kenya in 2012, since then the disease has rapidly spread to most parts of eastern and central Africa region including Tanzania, Burundi, DRC Congo, Rwanda, Uganda, Ethiopia and similar symptoms were observed in South Sudan. Elsewhere, the disease was caused by infection of <i>Maize Chlorotic Mottle Virus</i> (MCMV) in combination with any of the potyviruses namely; <i>maize dwarf mosaic virus</i> (MDMV), <i>sugarcane mosaic virus</i> (SCMV) and tritimovirus <i>wheat streak mosaic virus</i> (WSMV). In Africa, the disease occurs due to combined infections of maize by MCMV and SCMV, leading to severe yield losses. Efforts to address the disease spread have been ongoing. Serological techniques including enzyme-linked immuno-sorbent assay (ELISA), polymerase chain reaction (PCR), genome-wide association (GWAS) mapping and next generation sequencing have been effectively used to detect and characterize MLN causative pathogens. Various management strategies have been adapted to control MLN including use of resistant varieties, phytosanitary measures and better cultural practices. This review looks at the current knowledge on MLN causative viruses, genetic architecture and molecular basis underlying their synergistic interactions. Lastly, some research gaps towards MLN management will be identified. The information gathered may be useful for developing strategies towards future MLN management and maize improvement in Africa.</p>","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7753892/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38766408","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_3
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Viral Proliferation and Replication","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_3","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_3","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"89 1 1","pages":"31 - 38"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79424571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_13
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Laboratory Methods for Detecting Viral Infections","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_13","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_13","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"18 1","pages":"163 - 181"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75897841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_14
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Viruses with Single-Stranded, Positive-Sense RNA Genomes","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_14","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_14","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"119 1","pages":"185 - 349"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84897256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_2
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Viruses: Definition, Structure, Classification","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_2","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_2","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"165 1","pages":"17 - 30"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85000516","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_1
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Historical Overview","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_1","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_1","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"36 1","pages":"3 - 15"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85017673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_6
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Transformation and Carcinogenesis","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_6","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_6","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"14 1","pages":"57 - 67"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89805180","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_12
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Viral Evolution","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_12","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_12","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"2 1","pages":"155 - 161"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76630036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-12DOI: 10.1007/978-3-642-20718-1_10
S. Modrow, D. Falke, U. Truyen, H. Schätzl
{"title":"Vaccines","authors":"S. Modrow, D. Falke, U. Truyen, H. Schätzl","doi":"10.1007/978-3-642-20718-1_10","DOIUrl":"https://doi.org/10.1007/978-3-642-20718-1_10","url":null,"abstract":"","PeriodicalId":93204,"journal":{"name":"Journal of general and molecular virology","volume":"7 1","pages":"135 - 146"},"PeriodicalIF":0.0,"publicationDate":"2013-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87597695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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