Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus

Li, Qiong; Yue, Zhiqin; Liu, Hong; Liang, Chengzhu; Zheng, Xiaobo; Zhao, Yuqing; Chen, Xiao; Xiao, Xia; Chen, Changfu

doi:10.1016/j.jviromet.2009.10.028

Cited by 33 publications

(27 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…LAMP is more specific than PCR, because 4 or 6 primers used in LAMP increase specificity (45). Although short detection time and specificity of LAMP are the major advantages, continuous amplification of the target gene by LAMP may be confused with nonspecific PCR products on gel electrophoresis.…”

Section: Discussionmentioning

confidence: 99%

“…Although short detection time and specificity of LAMP are the major advantages, continuous amplification of the target gene by LAMP may be confused with nonspecific PCR products on gel electrophoresis. In order to overcome this disadvantage of LAMP, alternative LAMP-based detection techniques were recently proposed (45)(46)(47)(48). In one variation that does not use gel electrophoresis, a positive reaction was determined by Ca 2ϩ precipitation and SYBR green fluorescent or color dye with naked-eye inspection (45,47).…”

Section: Discussionmentioning

confidence: 99%

“…In order to overcome this disadvantage of LAMP, alternative LAMP-based detection techniques were recently proposed (45)(46)(47)(48). In one variation that does not use gel electrophoresis, a positive reaction was determined by Ca 2ϩ precipitation and SYBR green fluorescent or color dye with naked-eye inspection (45,47). In addition, real-time LAMP and probe-based LAMP have also been reported (46,48).…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

et al. 2014

View full text Add to dashboard Cite

This study aimed to develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Arcobacter species. Specific primers targeting the 23S ribosomal RNA gene were used to detect Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The specificity of the LAMP primer set was assessed using DNA samples from a panel of Arcobacter and Campylobacter species, and the sensitivity was determined using serial dilutions of Arcobacter species cultures. LAMP showed a 10-to 1,000-fold-higher sensitivity than multiplex PCR, with a detection limit of 2 to 20 CFU per reaction in vitro. Whereas multiplex PCR showed cross-reactivity with Campylobacter species, the LAMP method developed in this study was more sensitive and reliable than conventional PCR or multiplex PCR for the detection of Arcobacter species.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Loop-mediated isothermal amplification (LAMP) is a technique that has been developed to amplify nucleic acids with high specificity, sensitivity, and rapidity under isothermal conditions [8]. In aquaculture, LAMP assays have been developed to detect fish and shellfish pathogens including aquatic DNA viruses such as red seabream iridovirus (RSIV) [9], white spot syndrome virus (WSSV) [10–14], koi herpesvirus (KHV, CyHV-3) [15–19], infectious hypodermal and hematopoietic necrosis virus (IHHNV) [20–22], hepatopancreatic parvovirus (PmDNV) [23], Singapore grouper iridovirus (SGIV) [24], acute viral necrobiotic virus (AVNV) [25], turbot reddish body iridovirus (TRBIV) [26], infectious spleen and kidney necrosis virus (ISKNV) [27], lymphocystis disease virus (LCDV) [28], ostreid herpesvirus 1 (OsHV-1) [29], and soft shelled turtle iridovirus (STIV) [30]. Therefore, LAMP has been a useful tool for the diagnosis of aquatic animal diseases.…”

Section: Introductionmentioning

confidence: 99%

A Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cyprinid Herpesvirus 2 in Gibel Carp (Carassius auratus gibelio)

Zhang

Zeng

Fan

et al. 2014

The Scientific World Journal

View full text Add to dashboard Cite

A rapid and sensitive loop-mediated isothermal amplification (LAMP) assay for Cyprinid herpesvirus 2 (CyHV-2) detection in gibel carp was developed. Following cloning and sequencing of the putative DNA helicase gene of CyHV-2 isolate from China, a set of four specific primers was designed based on the sequence. The MgCl2 concentration and the reaction temperature were optimized to 6 mM, 64°C, respectively. LAMP products were detected by visual inspection of a color change due to addition of SYBR Green I stain. The specificity and sensitivity of the LAMP assay were determined. No cross-reaction was observed with other fish DNA viruses including eel herpesvirus, koi herpesvirus, and Chinese giant salamander iridovirus. The LAMP assay was found to be equally sensitive as nested PCR. A comparative evaluation of 10 fish samples using LAMP and nested PCR assays showed an overall correlation in positive and negative results for CyHV-2. These results indicate that the LAMP assay is simple, sensitive, and specific and has a great potential use for CyHV-2 detection in the laboratory and field.

show abstract

“…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

View full text Add to dashboard Cite

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.

show abstract

Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus

Cited by 33 publications

References 23 publications

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

A Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cyprinid Herpesvirus 2 in Gibel Carp (Carassius auratus gibelio)

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

Contact Info

Product

Resources

About