Detection of Equine Rotavirus by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP)

Nemoto, Manabu; Imagawa, Hiroshi; Tsujimura, Koji; Yamanaka, Takashi; Kondo, Takashi; Matsumura, Tomio

doi:10.1292/jvms.09-0446

Cited by 33 publications

(32 citation statements)

References 19 publications

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“…Sci. 72(9): 1247-1250, 2010 Equine rotavirus is a non-enveloped virus belonging to the genus Rotavirus in the family Reoviridae. Equine rotavirus infection is a major cause of diarrhea in foals up to 3 months of age [3,10].…”

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confidence: 99%

“…The RT-PCR products were analyzed by gel electrophoresis. The RT-LAMP assay was performed with the primers designed to target the VP4 gene of the genotype P [12] strain [15]. The RT-LAMP reaction mixture was prepared using a Loopamp ® RNA Amplification Kit (Eiken Chemical Co. Ltd., Tokyo, Japan) and fluorescent detection reagent (Eiken Chemical Co. Ltd.) in accordance with the manufacturer's instructions.…”

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confidence: 99%

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Evaluation of Rapid Antigen Detection Kits for Diagnosis of Equine Rotavirus Infection

Nemoto

Hata

Higuchi

et al. 2010

The Journal of Veterinary Medical Science

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ABSTRACT. We evaluated antigen detection kits for human rotavirus with regard to their usefulness for diagnosing equine rotavirus infection. Limiting dilution analyses showed that of the seven kits investigated the Dipstick 'Eiken' Rota (Dipstick) had the highest sensitivity to two serotypes of equine rotavirus. The Dipstick did not cross-react with several equine intestinal pathogens. An investigation using 249 fecal samples indicated that the sensitivity of the Dipstick was 81.9% and 47.3%, and its specificity was 98.2% and 99.0%, and its concordance rate was 92.8% and 68.3%, compared with values obtained using reverse transcription polymerase chain reaction and reverse transcription loop-mediated isothermal amplification, respectively. Although a negative result does not preclude the possibility of equine rotavirus infection, the Dipstick would be useful as routine test for diagnosing equine rotavirus infection in daily clinical practice because of its ease of handling.KEY WORDS: diagnosis, equine rotavirus, rapid antigen detection.J. Vet. Med. Sci. 72(9): 1247-1250, 2010 Equine rotavirus is a non-enveloped virus belonging to the genus Rotavirus in the family Reoviridae. Equine rotavirus infection is a major cause of diarrhea in foals up to 3 months of age [3,10]. Equine rotavirus has two outer capsid proteins, VP7 and VP4, which are independent neutralization antigens. VP7 and VP4 are classified as G (glycoprotein) serotypes by virus neutralization tests and P (protease-sensitive) genotypes by genetic analyses [14], respectively. Only G3 P[12] and G14 P[12] strains are currently considered to be circulating in the horse populations of Japan, Australia, Germany, Ireland and the United Kingdom [2,4,7,13,16]. Equine rotavirus infection has been diagnosed by electron microscopy, virus isolation, serological tests and molecular diagnostic methods [1,5]. These methods are not routinely available in diagnostic laboratories because they require sophisticated techniques and expensive equipment. It is difficult for clinical veterinarians to conduct these diagnostic methods as part of their daily clinical practice. Therefore, we focused our attention on rapid antigen detection (RAD) kits for use in routine tests in daily clinical practice. RAD kits do not require additional reagents, equipment or expertise, and results can be obtained within 15 min. RAD kits are designed to detect human group A rotaviruses based on the principle of latex agglutination assay (LA) or immunochromatographic assay. Because equine rotavirus is classified as a group A rotavirus, it is anticipated that RAD kits would also work for the rapid diagnosis of equine rotavirus infection. In fact, one LA kit (Virogen Rotatest, Wampole Laboratories, N.J., U.S.A.), which is not commercially available in Japan, has been reported to be useful for diagnosing equine rotavirus infection [6]. Although several RAD kits are available in Japan, their performance as regards the diagnosis of equine rotavirus infection has not been examined. In this study, we have ...

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Evaluation of Rapid Antigen Detection Kits for Diagnosis of Equine Rotavirus Infection

Nemoto

Hata

Higuchi

et al. 2010

The Journal of Veterinary Medical Science

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“…This finding suggests that in future strangles outbreaks, use of the seM-LAMP developed in this study will benefit the response by reducing detection time. Moreover, the seM-LAMP can produce similar results using a kit for visual analysis of turbidity and an incubator to maintain constant temperature even if the real-time turbidimeter used in this study is not used [9]. Based on this, the potential for applying the seM-LMAP in clinical situations seems promising.…”

Section: Discussionmentioning

confidence: 84%

“…The loop-mediated isothermal amplification (LAMP) method was developed as a nucleic acid amplification method to replace the PCR method, and the LAMP method has been clinically applied as a rapid diagnostic method for various pathogens in recent years [9,10,12,17,18]. LAMP amplifies a nucleic acid with high specificity, efficiency, and speed under isothermal conditions.…”

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Development and Application of Loop-Mediated Isothermal Amplification Methods Targeting the seM Gene for Detection of Streptococcus equi subsp. equi

Hobo

Niwa

Oku

2012

The Journal of Veterinary Medical Science

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ABSTRACT. Loop-mediated isothermal amplification (LAMP) constitutes a potentially valuable diagnostic tool for rapid diagnosis of contagious diseases. In this study, we developed a novel LAMP method (seM-LAMP) to detect the seM gene of Streptococcus equi subsp. equi (S. equi), the causative agent of strangles in equids. The seM-LAMP successfully amplified the target sequence of the seM gene at 63C within 60 min. The sensitivity of the seM-LAMP was slightly lower than the 2nd reaction of the seM semi-nested PCR. To evaluate the species specificity of the seM-LAMP, we tested 100 S. equi and 189 non-S. equi strains. Significant amplification of the DNA originating from S. equi was observed within 60 min incubation, but no amplification of non-S. equi DNA occurred. The results were identical to those of seM semi-nested PCR. To investigate the clinical usefulness of the methods, the seM-LAMP and the seM semi-nested PCR were used to screen 590 nasal swabs obtained during an outbreak of strangles. Both methods showed that 79 and 511 swabs were S. equi positive and negative, respectively, and the results were identical to those of the culture examination. These results indicate that the seM-LAMP is potentially useful for the reliable routine diagnosis of Streptococcus equi subsp. equi infections.

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“…Laboratory diagnosis of HSV-1 is an important public health tool, given the proven ability of this viral infection to cause enormous morbidity and financial burden particularly in immune compromised individuals (7,8). HSV-1 causes devastating infections of newborns, including fatal encephalitis or inflammation of the brain (8,9).…”

Section: Discussionmentioning

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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Detection of Equine Rotavirus by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP)

Cited by 33 publications

References 19 publications

Evaluation of Rapid Antigen Detection Kits for Diagnosis of Equine Rotavirus Infection

Evaluation of Rapid Antigen Detection Kits for Diagnosis of Equine Rotavirus Infection

Development and Application of Loop-Mediated Isothermal Amplification Methods Targeting the <i>seM</i> Gene for Detection of <i>Streptococcus equi</i> subsp. <i>equi</i>

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

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