Development of a loop-mediated isothermal amplification for rapid detection of orf virus

Tsai, Su-Ming; Chan, Kuei-Chuan; Hsu, Wei‐Li; Chang, Tien‐Jye; Wong, Min‐Liang; Wang, Chi-Young

doi:10.1016/j.jviromet.2009.01.003

Cited by 74 publications

(62 citation statements)

References 16 publications

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“…One may conclude that RT-LAMP works faster (4-fold) with higher sensitivity (40-fold) for detecting of PLRV compared with RT-PCR. These results parallel with previous studies, suggesting that RT-LAMP can be 10 to 100-fold more sensitive than RT-PCR [24,[32][33][34]. Overall, our results showed that some ingredients of RT-LAMP reaction, such as MgSO 4 , Betaine, primers, dNTPs, and DNA polymerase concentrations, as well as temperature and time period of the reaction play important roles in efficiency of the assay.…”

Section: It Is a Rapid And A Highly Specific Methodssupporting

confidence: 91%

“…However, the turbidity of the positive samples is not stable for a longer time and it should be judged soon after taking out of the samples from the water bath or from the thermal cycler. As a solution, adding SYBR Green and Ethidium Bromide to the tubes will provide enough time to monitor changes in the color of the tubes and to detect the positive samples under UV illumination as stressed by other workers [25,33]. However, using of toxic staining materials and UV irradiation would not be compatible with the main feature of this technique which is "safety".…”

Section: It Is a Rapid And A Highly Specific Methodsmentioning

confidence: 99%

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Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Almasi¹

2012

J Plant Pathol Microb

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Loop-mediated isothermal amplification assay amplifies DNA/RNA with high specificity and sensitivity. In this study, we describe an optimized reverse transcription-LAMP assay for detection of Potato Leafroll Virus. Firstly, DAS-ELISA assay was performed to detect of the virus in a collection containing 40 suspicious samples. Lastly, two samples were detected as the positive samples. Then, the positive samples were verified by RT-PCR and RT-LAMP methods. Furthermore, the results demonstrated that the RT-LAMP assay was 40 times sensitive and 4 time faster compared to RT-PCR. RT-LAMP assay was accomplished in the water bath either frees from any thermal cycler machine or sophisticated laboratories facility. Moreover, in RT-LAMP reaction the positive samples were detected through turbidity which produced by magnesium pyrophosphate. Interestingly, the application of CaCl 2 instead of MgSO 4 which create calcium pyrophosphate in reaction could significantly increase both stability and concentration of turbidity. Consequently, it could be an interesting alternative to MgSO4. Overall, the newly developed RT-LAMP assay can be a sensitive, specific and low-cost method for early detection of Potato Leafroll Virus and also other viral plant pathogens.

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Section: It Is a Rapid And A Highly Specific Methodssupporting

confidence: 91%

Section: It Is a Rapid And A Highly Specific Methodsmentioning

confidence: 99%

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Almasi¹

2012

J Plant Pathol Microb

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“…Li et al (2013b) developed a method for rapid detection of orf virus by LAMP assay based on DNA polymerase gene. LAMP assay is an easy, quick, precise and sensitive technique for detection of orf virus infection without any cross reactivity with other viruses (Tsai et al, 2009;Wang et al, 2013;.…”

Section: Advances In Animal and Veterinary Sciencesmentioning

confidence: 99%

Contagious Pustular Dermatitis (Orf Disease) - Epidemiology, Diagnosis, Control and Public Health Concerns

Kumar¹,

Trivedi²,

Bhatt³

et al. 2015

Adv. Anim. Vet. Sci.

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| Contagious pustular dermatitis (CPD), also known as Orf or contagious ecthyma is an important viral disease of sheep and goats. It is mainly seen as a benign disease but malignant form has also been reported from few parts of the world. The rates of morbidity and mortality are higher, particularly in lambs and kids experiencing the disease for the first time. The causative agent of disease is Orf virus, type species of the genus Parapoxvirus belonging to family Poxviridae. The virus produces localized persistent proliferative skin lesions and affected hosts are infected repeatedly owing to its host immune evasive strategy. These viruses have been found uniformly labile to chloroform but resistant to ether and also exhibit antigenic variations among them. Clinically, the disease is enzootic and occurs in three forms viz. labial, genital / mammary and generalized forms. The incubation period in natural cases is 2-3 weeks. The disease outbreaks mostly occur between autumn and spring but its severity is more in autumn and winter than spring. The virus grows well in cell cultures of caprine, ovine and bovine origin. Confounding symptoms impose laboratory affirmation by serological assays or molecular techniques. The disease can be diagnosed by electron microscopy, serological tests and PCR/ quantitative real-time PCR. Virus specific antibody response to structural proteins of capripox and orf viruses in western-blot analysis readily differentiates these two infections. The antibody response to the 32 kDa and 26 kDa proteins of capripox viruses provides a firm basis for differentiation. Although, the role of humoral immunity is well established but probably cell mediated immunity plays a major role in recovery from natural infections. A number of inactivated and live or live modified vaccines have been tried with variable success. The duration of immunity after vaccination is controversial; outbreaks have occurred in vaccinated animals. The disease is also of public health significance as it causes infection in human beings. Keywords

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“…Notomi et al (2000) first described an autocycling strand displacement DNA synthesis using the Bst DNA polymerase, the LAMP method is carried out widely in diagnostic clinical laboratories for detection of bacteria and viruses (12). The LAMP assay as a rapid and cost-effective method does not require thermocycler and gel electrophoresis in some cases (using turbidity) and the inhibitory effects of substances in samples are largely eliminated (4,13). A reliable LAMP mostly depends on the specificity of the primer sets (4).…”

Section: Loop-mediated Isothermal Amplification (Lamp) For …mentioning

confidence: 99%

“…In this method, four primers (two inner primers and two outer primers) plus two loop primers that generally, recognize six to eight regions of the target DNA are used which makes the test highly sensitive (3). In addition, the thermal cycler is not necessary and reaction is performed in a water bath or heat block under isothermal conditions (4,5). In the last decade, LAMP assay as a rapid, efficient and cost-effective method which has been set up to detect different kind of viral infection is considered to be use for the diagnosis of viral infection, and for prevention of viral outbreaks.…”

Section: Introductionmentioning

confidence: 99%

Loop-Mediated Isothermal Amplification (LAMP) for the Rapid Diagnosis of Herpes Simplex Virus Type 1 (HSV-1)

Pourhossein

Soleimanjahi

Behzadian³

et al. 2011

Iran J Virol

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Background and Aims: considering difficulties in usual laboratory methods in detection of viral infections, improved DNA-based diagnostic techniques are more reliable. Loop mediated isothermal amplification method (LAMP) is a nucleic acid amplification method that amplifies DNA using six primers which has been developed to diagnose viruses as a rapid and high efficiency test. In this study, the LAMP was used for detection of Herpes simplex virus. Methods: The set of primers were designed, for detection of HSV-based on gG fragment. The genome of the virus was extracted from the culture supernatant of infected cells. LAMP technique was optimized in term of temperature, time and the ingredients used for test. For detection of HSV-1 infection, sensitivity and specificity of this technique were determined by various dilutions of virus and infected samples with HSV-2 that was taken from Day Hospital of Tehran. Results: The sensitivity and specificity of HSV-1 specific LAMP method, reached about 500 copies/tube and 99.9% respectively. Furthermore, both the agarose gel electrophoresis containing Ethidium Bromide (EtBr) and the turbidity assay directly detected HSV-1 virus LAMP products in reactions. Conclusion: In this study, the reliability of LAMP for detection of HSV-was approved; therefore this rapid, accurate, and cost-effective detection and quantification method may perhaps be an investigative tool which can be valid for detection of target viral genome in clinical specimens in the absence of the necessary facilities for performing PCR.

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Development of a loop-mediated isothermal amplification for rapid detection of orf virus

Cited by 74 publications

References 16 publications

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Detection of Coat Protein Gene of the Potato Leafroll Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

Contagious Pustular Dermatitis (Orf Disease) - Epidemiology, Diagnosis, Control and Public Health Concerns

Loop-Mediated Isothermal Amplification (LAMP) for the Rapid Diagnosis of Herpes Simplex Virus Type 1 (HSV-1)

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