Detection of PCV2 DNA by SYBR Green I-based quantitative PCR

Yang, Zong-zhao; Habib, Md. Ahasan; Shuai, Jiangbing; Fang, Weihuan

doi:10.1631/jzus.2007.b0162

Cited by 40 publications

(34 citation statements)

References 21 publications

(32 reference statements)

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“…Detection over a wide dynamic range of concentrations by ddPCR allows for the measurement of viral ranges in animals, which can exhibit various levels of infection. 16 In the present study, the linear regression correlation coefficients were measured using either ddPCR or TaqMan real-time PCR showed good R 2 values that were ~1 (Fig. 2).…”

supporting

confidence: 60%

Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

et al. 2015

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Abstract. Sensitive detection of Porcine circovirus-2 (PCV-2) is very important for surveillance of postweaning multisystemic wasting syndrome. Droplet digital polymerase chain reaction (ddPCR) is novel PCR method that can achieve high precision. Our study aimed to develop a sensitive assay utilizing ddPCR to detect PCV-2. Specificity of the assay was confirmed by the failure of amplification of DNA of other relevant viruses. The detection limit for ddPCR was 25 copies/μL, a 4-fold greater sensitivity than TaqMan real-time PCR. Both methods showed a high degree of linearity (R 2 = ~1), although TaqMan real-time PCR showed less sensitivity than ddPCR for clinical detection. Our findings indicate that ddPCR might represent a promising platform for detecting PCV-2 viral loads.

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supporting

confidence: 60%

Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

et al. 2015

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“…In contrary, Ct of NTC seems increase as lower annealing temperature was used (Figure 3ac). Presence of NTC was common, especially when non specific dye such as SYBR green I was used (Yang et al, 2007). Bustin et al, (2009) reported that NTC was negligible at distance of Ct sample was more than 5 because, in this distance, Ct of NTC and sample was distinguishable.…”

Section: Optimization Of Tm Shift Sybr Green I Qpcr Techniquementioning

confidence: 99%

Method Development for Detection of E545A Mutation PIK3CA Gene in Breast Cancer Patients Using Tm Shift SYBR Green I qPCR

Ahwani

Desriani

Widyastuti

et al. 2017

Indones. J. Biotechnol.

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E545A is one of the point mutations, and its frequency is high in PIK3CA gene (3.8%), particularly breast cancer patients in Singapore (13.8%) and Mexico (11.5%). In addition to induce breast cancer, the mutation also caused resistance of antiHER2 in HER2 cancer subtype. The tremendous effect of this mutation was not supported by affordable detection method. This study aimed to develop a feasible and sensitive method for E545A detection. The developing method used Tm and Ct to identify samples. Based on optimization, the best condition was obtained with annealing temperature of 65°C. At this condition, Tm and Ct of each sample were (a) exon 9 (78.4°C and 13.005±0.007) and (b) E5454A (80.4°C and 10.07±0.1). This method also demonstrated good precision as observed in coefficient variance of intra and inter assay (0). Thus, method for E5454A detection mutation was successfully developed.

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“…We used PCR to detect Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Pasteurella multocida, porcine circovirus 2 (PCV2), Suid herpesvirus 1 (SuHV-1), and porcine parvovirus (PPV) as described previously (Artiushin et al 1993;Register and DeJong 2006;McKillen et al 2007;Yang et al 2007;Tang et al 2009). The extracted DNA was also tested for b-actin as described by Hui et al (2004).…”

Section: Detection Of Nucleic Acids From Different Infectious Agentsmentioning

confidence: 99%

SWINE INFECTIOUS AGENTS INTAYASSU PECARIANDPECARI TAJACUTISSUE SAMPLES FROM BRAZIL

Castro

Brombila

Bersano

et al. 2014

Journal of Wildlife Diseases

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ABSTRACT:Peccaries and pigs, Tayassuidae and Suidae respectively, diverged approximately one million years ago from a common ancestor. Because these families share some pathogens, peccaries can act as reservoirs of infectious pathogens for domestic and wild swine. We evaluated the presence of swine infectious agents in the spleen and lung tissues of white-lipped peccaries (WLP; Tayassu pecari) and collared peccaries (CP; Pecari tajacu) in Brazil. Samples from 10 adult CP and three WLP, which had been hunted by locals or hit by motor vehicles, were obtained from two free-ranging Brazilian populations. The samples were tested by PCR for Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Pasteurella multocida, porcine circovirus 2 (PCV2), Suid herpesvirus 1 (SuHV-1), and porcine parvovirus (PPV). Positive samples were sequenced. Both species were negative for PPV and B. bronchiseptica and positive for PCV2 and SuHV-1. The lungs of two animals were positive for M. hyopneumoniae and P. multocida. This report is the first demonstration of PCV2 and SuHV-1 swine viruses and of M. hyopneumoniae and P. multocida bacteria in peccaries. One factor contributing to this detection was access to tissue samples, which is uncommon. The role of these infectious agents in peccaries is unknown and further epidemiologic studies should be performed. This study identified several infectious agents in peccaries and highlighted the importance of the tissue type used to detect pathogens.

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Detection of PCV2 DNA by SYBR Green I-based quantitative PCR

Cited by 40 publications

References 21 publications

Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

Method Development for Detection of E545A Mutation PIK3CA Gene in Breast Cancer Patients Using Tm Shift SYBR Green I qPCR

SWINE INFECTIOUS AGENTS INTAYASSU PECARIANDPECARI TAJACUTISSUE SAMPLES FROM BRAZIL

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