Development of TaqMan® MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1

Guo, Yufei; Cheng, Anchun; Wang, Mingshu; Shen, Cong; Jia, Renyong; Chen, Shun Le; Zhang, Na

doi:10.1186/1743-422x-6-71

Cited by 39 publications

(39 citation statements)

References 25 publications

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“…As prevention and early detection are the most logical strategies for pathogen control, the most effective way to control disease is by routinely screening for pathogens (Guo et al. 2009).…”

Section: Discussionmentioning

confidence: 99%

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop-mediated isothermal amplification

Yang

et al. 2010

Journal of Applied Microbiology

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Aim: The objective of this study is to develop a serovar‐specific loop‐mediated isothermal amplification (LAMP) method for sensitive, rapid, and inexpensive detection of Salmonella serovar Enteritidis under field conditions. Methods: A set of six specific primers was designed with Salmonella Enteritidis DNA as the target. LAMP conditions were optimized by incubating the target DNA with the Bst DNA polymerase large fragment in a simple water bath. The sensitivity and specificity of LAMP was then compared with those of fluorescent quantitative real‐time polymerase chain reaction (FQ‐PCR). Results: The results were as follows. (1) Serovar‐specific Salmonella Enteritidis DNA was amplified at 65°C in as early as 20 min in a water bath. (2) A colour change visible to the naked eye indicated a positive amplification reaction. (3) The detection limit of the LAMP assay was 4 copies μl−1; thus, the sensitivity and specificity of this assay is similar to those of the FQ‐PCR. Conclusions: LAMP is a high‐throughput detection technique with high sensitivity, specificity, and simplicity; these factors make it suitable for specifically detecting Salmonella Enteritidis under field conditions and in laboratory settings. Thus, LAMP eliminates the need for complicated equipment and technical training in the detection of this specific serovar. Significance and impact of the study: This is the first study involving the use of LAMP to detect Salmonella serovar‐specific DNA sequences. It is also the first to report an ideal method of distinguishing between Salmonella Enteritidis and other Salmonella under field conditions.

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Section: Discussionmentioning

confidence: 99%

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop-mediated isothermal amplification

Yang

et al. 2010

Journal of Applied Microbiology

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“…Since prevention and early detection are the most logical strategies for pathogen control, the most effective method of disease control is routine screening for pathogens [33]. Sensitive and rapid methods are required for the detection of PCMV under field conditions.…”

Section: Discussionmentioning

confidence: 99%

Development and evaluation of a loop-mediated isothermal amplification assay for the rapid detection of porcine cytomegalovirus under field conditions

Yang

Zhang

Liu

et al. 2012

Virol J

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BackgroundPorcine cytomegalovirus (PCMV) induces silent infection in adult pigs but more frequently causes fatal, generalized infection in newborn piglets. This study aimed to develop a new loop-mediated isothermal amplification (LAMP) method for the sensitive, rapid, and inexpensive detection of PCMV under field conditions.MethodsTissue obtained from nine-week-old PCMV-free Landrace pigs or pig samples from postmortem examinations were analyzed. The samples were found to have clinical signs and lesions consistent with inclusion body rhinitis. Six specific primers were designed by targeting the PCMV DNA polymerase (DPOL) DNA. The LAMP reaction was optimized in a water bath. The sensitivity and specificity of LAMP and polymerase chain reaction (PCR) were compared.ResultsPCMV DNA was amplified at 65°C, and the result could be detected as early as 30 min into the reaction. Positive reactions could be visualized by the naked eye as a color change brought on by the addition of SYBR Green. The sensitivity and specificity of LAMP were found to be similar to those of the PCR.ConclusionsLAMP is a high-throughput technique for the detection of PCMV and has a high specificity, sensitivity and simplicity; these factors make it suitable for detection of PCMV under field conditions.

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“…The VP1 gene was amplified using conventional PCR (primers F1 and R1) with Premix Taq (Takara Bio Inc., Shiga, Japan). The purified PCR product was cloned into the pGM-Simple-T Fast vector (Tiangen Corp., Beijing, China) and transformed into Escherichia coli DH5α competent cells [21]. The recombinant plasmid named pVP1 was extracted using the TIANprep plasmid extraction kit (Tiangen Corp.) and sequenced.…”

Section: Methodsmentioning

confidence: 99%

Development of a SYBR Green II Real-Time Polymerase Chain Reaction for the Clinical Detection of the Duck-Origin Goose Parvovirus in China

Luo

Chen

et al. 2018

Intervirology

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Objective: To establish an efficient, convenient and quantitative method for the clinical detection of the duck-origin goose parvovirus. Method: In the present study, a real-time polymerase chain reaction (PCR) method was established for detecting the duck-origin goose parvovirus using the fluorescent chimeric dye SYBR Green II. Specific primers were designed to target a highly conserved region of the VP3 gene of the duck-origin goose parvovirus. Results: This method was able to detect a minimum of 19.6 copies/μL of viral genomic DNA. Results showed that this method was faster and had a higher sensitivity than the traditional PCR in the clinical specimen test. In this paper, we developed a rapid, sensitive detection and quantitative analysis technology for the duck-origin goose parvovirus by real-time PCR assay. Conclusion: This test provides improved technical support for studies regarding the clinical diagnosis and epidemiological investigations of the duck-origin goose parvovirus.

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Development of TaqMan® MGB fluorescent real-time PCR assay for the detection of anatid herpesvirus 1

Cited by 39 publications

References 25 publications

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop-mediated isothermal amplification

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop-mediated isothermal amplification

Development and evaluation of a loop-mediated isothermal amplification assay for the rapid detection of porcine cytomegalovirus under field conditions

Development of a SYBR Green II Real-Time Polymerase Chain Reaction for the Clinical Detection of the Duck-Origin Goose Parvovirus in China

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