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ارزیابی آزمون تکثیر همدمای وابسته به حلقه در شناسایی ویروس موزاییک کاهو (LMV) | ||
| فصلنامه علمی زیست فناوری گیاهان زراعی | ||
| مقاله 5، دوره 6، شماره 3 - شماره پیاپی 19، آبان 1396، صفحه 51-63 اصل مقاله (953.9 K) | ||
| نوع مقاله: علمی پژوهشی | ||
| نویسندگان | ||
| محمد امین الماسی1؛ گلاویژ الماسی* 2 | ||
| 1دانشگاه آزاد اسلامی، واحد تهران شمال، باشگاه پژوهشگران جوان و نخبگان، تهران، ایران | ||
| 2دانشگاه آزاد اسلامی، واحد تهران شمال، باشگاه پژوهشگران جوان و نخبگان، تهران، ایران. | ||
| چکیده | ||
| شایعترین ویروس تأثیرگذار بر کاهو در سراسر مزارع جهان، ویروس موزاییک کاهو (Lettuce mosaic virus; LMV) متعلق به خانواده Potyviridea و جنس Potyvirus است. روشهای متعددی شامل آزمونهای سرولوژیکی و مولکولی جهت تشخیص این ویروس وجود دارند، اما این روشها زمانبر بوده و نیازمند ابزارهای پیچیدهای هستند. هدف از این پژوهش، کاهش زمان موردنیاز جهت تشخیص LMV با استفاده از روش تکثیر همدمای وابسته به حلقه (LAMP) بود. نمونههای برگی (38 نمونه) با علایم مشابه به ویروس موزاییک کاهو از سطح استان کردستان جمعآوری و برای آزمون سرولوژیکی (DAS-ELISA) بهکارگرفته شدند و در نهایت چهار نمونه مثبت تشخیص داده شد. سپس RNA کل استخراج و واکنش رونویسی معکوس LAMP به صورت یک مرحلهای، تحت شرایط همدما انجام شد و نتایج به وسیله الکتروفورز بر روی ژل آگارز و رنگ هیدروکسینفتولبلو (HNB) ارزیابی شد. نتیجه مثبت استفاده از رنگ HNB تغییر رنگ مخلوط واکنش از بنفش به آبی آسمانی بود. علاوه بر این، در اینجا آزمون جدید IC-RT-LAMPجهت تشخیص سریع و آسانLMV (بدون نیاز به استخراج RNA) توسعه داده شد و کارآیی آن با آزمونهای دیگر مقایسه شد. از مزایای روش RT-LAMP در مقایسه با سایر روشهای پیشین میتوان به اختصاصیت بالا، حساسیت بالا، سرعت بالا، کارآیی بالا، ایمنی، سهولت، عدم نیاز به تجهیزات پرهزینه جهت تکثیر، عدم نیاز به استخراج RNA (در آزمون IC-RT-LAMP)، عدم نیاز به کارهای تشخیصی بعد از تکثیر، تشخیص مشاهدهای و کاربردوستی آن اشاره کرد. | ||
| کلیدواژهها | ||
| DAS-ELISA؛ IC-RT-LAMP؛ ویروس موزاییک کاهو؛ RT-LAMP | ||
| موضوعات | ||
| بیوتکنولوژی بیماریهای گیاهی | ||
| عنوان مقاله [English] | ||
| Evaluation of loop mediated isothermal amplification assay for identification of Lettuce Mosaic Virus (LMV) | ||
| نویسندگان [English] | ||
| Mohammad Amin Almasi1؛ Galavizh Almasi2 | ||
| 1Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran | ||
| 2Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran | ||
| چکیده [English] | ||
| The most common virus affecting lettuce (Lactuca sativa L.) in the field worldwide is Lettuce mosaic virus (LMV), belonging to the family Potyviridea, genius Potyvirus. There are several techniques to detect LMV including serological and molecular assays, but these methods take a long time and requiring sophisticated tools. The aim of this study was to reduce the time required to detect LMV, using loop-mediated isothermal amplification (LAMP) technique. Leaf samples (38 samples) with symptoms similar to LMV were collected from Kurdestan province and were subjected to a serological (DAS-ELISA) test and lastly, four samples were detected as the positive samples. Then, total RNA was extracted and one-step reverse transcription (RT)-LAMP was carried out under isothermal conditions and results were evaluated by agarose gel electrophoresis and hydroxynaphthol blue (HNB) dye. A positive result using the HNB dye was a color change in master-mix from the violet to sky blue. In addition, a novel immunocapture (IC)-RT-LAMP assay for rapid and easy detection (without RNA extraction) of LMV was developed here and its potential compared with other assays. The advantages of RT-LAMP method in compare to other methods include the high specificity, high sensitivity, high rapidity, high efficiency, safety, fascinatingly, no requirement of expensive and tools for amplification, no need to RNA extraction (in IC-RT-LAMP assay), no post-amplification treatment of the amplicons, visual detection and user friendly. | ||
| کلیدواژهها [English] | ||
| DAS- ELISA, IC-RT-LAMP, Lettuce mosaic virus, RT-LAMP | ||
| مراجع | ||
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Ahmadi S, Almasi MA, Fatehi F, Struik PC, Moradi A (2012) Visual detection of Potato leafroll virus by one-step reverse transcription loop-mediated isothermal amplification of DNA with hydroxynaphthol blue dye. J. Phytopathol. 161: 120-124.
Almasi MA (2015) Establishment and Application of a Reverse Transcription Loop-mediated Isothermal Amplification Assay for Detection of Grapevine fanleaf virus. Mol. Biol. 4: 5.
Almasi MA (2016) Tracking and identification of Alfalfa mosaic virus (AMV) by loop mediated isothermal amplification assay. Crop Biotech. 14: 73-84.
Almasi MA, Aghapour-Ojaghkandi M, Aghaei S (2013) Visual detection of Curly top virus by the colorimetric loop-mediated isothermal amplification. J. Plant Pathol. Microb. 4: 198.
Almasi MA, Aghapour-ojaghkandi M, Bagheri K, Ghazvini M, Hosseyni-dehabadi SM (2015) Comparison and evaluation of two diagnostic methods for detection of npt II and GUS genes in Nicotiana tabacum. Appl. Biochem. Biotechnol. 175: 3599-3616.
Almasi MA, Almasi G (2016) Development and evaluation of a reverse transcription loop-mediated isothermal amplification assay for detection of Beet necrotic yellow vein virus. Arch. Virol. doi:10.1007/s00705-016-3116-0
Almasi MA, Dehabadi SH (2013) Colorimetric immunocapture reverse transcription loop-mediated isothermal amplification assay for rapid detection of the Potato virus Y. J. Plant Pathol. Microb. 4: 188.
Almasi MA, Dehabadi SH, Eftekhari Z (2013) Immunocapture loop mediated isothermal amplification for rapid detection of Tomato yellow leaf curl virus (TYLCV) without DNA extraction. J. Plant Pathol. Microb. 4: 185.
Almasi MA, Erfanmanesh M, Jafary H, Hosseinidehabadi SM (2013) Visual detection of Potato leafroll virus by one-step reverse transcription loop-mediated isothermal amplification of DNA with the GenefinderTM dye. J. Virol. Methods 192: 51-54.
Almasi MA, Haghnazari A, Moradi A, Saberfar E (2012) Dِifferent methods of RT-LAMP for detection of potato leaf roll virus (PLRV). Gen. Engineer. Biosafety J. 1: 1-8.
Almasi MA, Hosseyni-Dehabadi SM, Aghapour-Ojaghkandi M (2014) Comparison and evalution of three diagnostic methods for detection of Beet curly top virus in sugar beet using different visualizing systems. Appl. Biochem. Biotechnol. 173: 1836-1848.
Almasi MA, Jafary H, Moradi A, Zand N, Ojaghkandi MA, Aghaei S (2013) Detection of coat protein gene of the Potato leafroll virus by reverse transcription loop-mediated isothermal amplification. J. Plant Pathol. Microb. 4: 156.
Almasi MA, Moradi A, Nasiri J, Karami S, Nasiri M (2012) Assessment of performance ability of three diagnostic methods for detection of Potato leafroll virus (PLRV) using different visualizing systems. Appl. Biochem. Biotechnol. 168: 770-784.
Almasi MA, Moradi A, Ojaghkandi MA, Aghaei S (2013) Development and application of loop-mediated isothermal amplification assay for rapid detection of Fusarium oxysporum f. sp. lycopersici. J. Plant Pathol. Microb. 4: 177.
Almasi MA, Ojaghkandi MA, Hemmatabadi A, Hamidi F, Aghaei S (2013) Development of colorimetric loop-mediated isothermal amplification assay for rapid detection of the Tomato yellow leaf curl virus. J. Plant Pathol. Microb. 4: 153.
Bannerot H, Boulidard L, Marrou JE, Duteil M (1969) Study of the inheritance of tolerance to Lettuce mosaic virus in the variety Gallega de Invierno. Ann. Phytopathol. 1: 219-226.
Bos L, Huijberts N, Cuperus C (1994) Further observation on variation of Lettuce mosaic virus in relation to lettuce (Lactuca sativa), and a discussion of resistance terminology. Eur. J. Plant Pathol. 100: 293-314.
Bruckart WL, Lorbeer JW (1975) Recent occurrences of cucumber mosaic, lettuce mosaic and broadbean wilt viruses in lettuce and celery fields in New York. Plant Dis. Reptr. 59: 203-206.
Candresse T, Lot H, German-Retana S, Krause-Sakate R, Thomas J, Souche S, Delaunay T, Lanneau M, Le Gall O (2007) Analysis of the serological variability of Lettuce mosaic virus using monoclonal antibodies and surface plasmon resonance technology. J. Gen. Virol. 88: 2605-2610.
Clark MF, Adams AN (1977) Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol. 34: 475-483.
Clewley JP, Arnold C (1977) The multiple alignment module of LASERGENE. Methods Mol. Bio. 70: 119-129.
Drews M, Schonhof I, Krumbein A (1997) Contents of minerals, vitamins and sugars in Iceberg lettuce (Lactuca sativa var capitat L.) grown in greenhouse, dependent on cultivar development stage. Gartenbauwissenschaft. 62: 65-72.
Fukuta S, Iida T, Mizukami Y, Ishida A, Ueda J, Kanbe M, Ishimoto Y (2003) Detection of Japanese yam mosaic virus by RT-LAMP. Arch. Virol. 148: 1713-1720.
Gonnella M, Conversa G, Santamarina P, Serio F (2003) Yield and quality of lettuce grown in floating system using different sowing density and plant spatial arrangements. Acta Hort. 614: 687-692.
Haghnazari A, Almasi MA, Hoseini SM (2011) LAMP and guide of primer design by PrimerExplorerV4 software. Azarkelk press, Zanjan, Iran.
Hosseini SF, Almasi MA, Kardi MT, Moghim S, Karbasizade V (2014) Molecular Detection of Clostridium Difficile in Patients with Diarrhea via LAMP Technique. J. Mazandaran Univ. Med. Sci. 24: 36-42.
Jagger ICA (1921) A transmissible mosaic disease of lettuce. J. Agric. Res. 20: 741-737.
Jeong J, Ju H, Noh J (2014) A review of detection methods for the plant viruses. Res. Plant Dis. 20: 173-181.
Krause-Sakate R, Le Gall O, Fakhfakh H, Peypelut M, Marrakchi M, Varveri C, Pavan MA, Souche S, Lot H, Zerbini FM, Candresse T (2002) Molecular and biological characterization of Lettuce mosaic virus isolates reveals a distinct and widespread type of resistance breaking isolate: LMV-Most. Phytopathology 92: 563-572.
Kristkova E, Dolezalova I, Lebeda A, Vinter V, Novotna A (2008) Description of morphological characters of lettuce (Lactuca sativa L.) genetic resources. Hort. Sci. 35: 113-129.
Moradi A, Almasi MA, Jafary H, Mercado-Blanco J (2014) A novel and rapid loop-mediated isothermal amplification assy for the specific detection of Verticillium dahliae. J. Appl. Microbiol. 116: 942-954.
Moradi A, Nasiri J, Abdollahi H, Almasi M (2012) Development and evaluation of a loopmediated isothermal amplification assay for detection of Erwinia amylovora based on chromosomal DNA. Eur. J. Plant Pathol. 133: 609-620.
Nagamine K, Hase T, Notomi T (2002) Accelerated reaction by loop mediated isothermal amplification using loop primers. Mol. Cell. Probes. 16: 223-229.
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28 :e63.
Pereira LS, Chaves ALR, Azevedo Filho JA, Colariccio A (2012) Lettuce mosaic virus indexation in seeds and seedlings of lettuce genotypes. Rev. bras. sementes 34: 628-635.
Peypelut M, Krause-Sakate R, Guiraud T, Pavan MA, Candresse T, Zerbini FM, Le Gall O (2004) Specific detection of lettuce mosaic virus isolates belonging to the "Most" type. J. Virol. Methods 121: 119-124.
Pink DAC, Kostova D, Walkey DGA (1992) Differentiation of pathotypes of Lettuce mosaic virus. Plant Pathol. 41: 5-12.
Redondo E, Krause-Sakate R, Yang SJ, Lot H, Le Gall O (2001) Lettuce mosaic virus pathogenicity determinants in susceptible and tolerant lettuce cultivars map to different regions of the viral genome. Mol. Plant Microbe. Interact. 14: 804-810.
Rowhani A, Stace-Smith R (1979) Purification and characterization of potato leafroll virus. Virology. 98: 45-54.
Ryder EJ (1970) Inheritance of resistance to common lettuce mosaic. J. Am. Soc. Hortic. Sci. 95: 378-379.
Sakate KR, Fakhfakh H, Peypelut M, Pavan MA, Zerbini FM, Marrakchi M, Candresse T, Le Gall O (2004) A naturally occurring recombinant isolate of Lettuce mosaic virus. Arch. Virol. 149: 191-197.
Sharma P, Jain RK, Saha S, Kalia P (2013) First report of Lettuce mosaic virus infecting Lactuca sativa in India. Plant Dis. 97: 849.
Soleimani P, Mossahebi GH, Koohi Habibi M (2011) Identification of some viruses causing mosaic on lettuce and characterization of Lettuce mosaic virus from Tehran Province in Iran. Afr. J. Agric. Res. 6: 3029-3035.
Suzuki R, Yoshikawa T, Ihira M, Enomoto Y, Inagaki S, Matsumoto K, Kato K, Kudo K, Kojima S, Asano S (2006) Development of the loop-mediated isothermal amplification method for rapid detection of cytomegalovirus DNA. J. Virol. Methods 132: 216-221.
Thompson JD, Gibson TJ, Higgins DG (2002) Multiple sequence alignment using ClustalW and ClustalX. Curr. Protoc. Bioinformatics. doi: 10.1002/0471250953.bi0203s00
Torrance L (1998) Developments in serological methods to detect and identify plant viruses. Plant Cell Tiss. Org. Cult. 52: 27-32.
Zerbini FMT, koike S, Gilbertson R (1995) Biological and molecular characterization of Lettuce mosaic potyvirus isolates from the Salinas Valley of California. Phytopathology 85: 746-752. | ||
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