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Number of results: 5
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Abstract

In this study, a SYBR Green-based real-time quantitative polymerase chain reaction (qPCR) assay was developed for rapid detection of porcine parvovirus (PPV) 6. Primer pairs targeting the conserved regions of PPV6 Capsid gene were designed. Sensitivity analyses revealed the lowest detection limit of the SYBR Green-based real-time PCR assay to be 47.8 copies/μL, which indicated it was 1000 times higher than that found in the conventional PCR investigations. This assay was specific and showed no cross-species amplification with other six porcine viruses. The assay demonstrated high repeatability and reproducibility; the intra- and inter-assay coefficients of variation were 0.79% and 0.42%, respectively. The positive detection rates of 180 clinical samples with SYBR Green-based real-time PCR and conventional PCR were 12.22% (22/180) and 4.44% (8/180), respectively. Our method is sensitive, specific, and reproducible. The use of SYBR Green-based real-time PCR may be suitable for the clinical detection and epidemiological investigation of PPV6.

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Authors and Affiliations

P. Sun
C.X. Bai
D. Zhang
J. Wang
K.K. Yang
B.Z. Cheng
Y.D. Li
Y. Wang
ORCID: ORCID
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Abstract

Composition effect on electro-optic (EO) properties of a LiNbO3 (LN) single-crystal has been investigated in a Li2O-content range of 47.0–49.95 mol%. Some non-congruent LN crystals with different Li2O-contents were prepared by performing Li-deficient or Li-rich vapour transport equilibration treatments on as-grown congruent LN crystals. Unclamped EO coefficients γ13 and γ33 of these samples were measured by a Mach–Zehnder interferometric method. The measurements show that in the Li-deficient regime both γ13 and γ33 increase by ∼8% as Li2O-content decreases from the congruent 48.6 mol% to the 47.0 mol% in the Li-deficient regime. The feature is desired for the EO application of the Li-deficient crystal. In the near-stoichiometric regime, both γ13 and γ33 reveal a non-monotonic dependence. As the Li2O-content increases from the 48.6 mol%, the EO coefficient decreases. Around Li2O-content 49.5 mol%, a minimum is reached. After that, the EO coefficient recovers slowly. At the stoichiometric composition, it recovers to a value close to that at the congruent point. Comparison shows that different crystal growth methods give rise to different defect structure features and hence different composition effects.

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Authors and Affiliations

J.-Q. Xu
W.-Y. Du
Q. Sun
W.-H. Wong
D.-Y. Yu
E.Y.-B. Pun
D.-L. Zhang
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Abstract

MDAP-2 is a new antibacterial peptide with a unique structure that was isolated from house- flies. However, its biological characteristics and antibacterial mechanisms against bacteria are still poorly understood. To study the biological characteristics, antibacterial activity, hemolytic activi- ty, cytotoxicity to mammalian cells, and the secondary structure of MDAP-2 were detected; the results showed that MDAP-2 displayed high antibacterial activity against all of the tested Gram-negative bacteria. MDAP-2 had lower hemolytic activity to rabbit red blood cells; only 3.4% hemolytic activity was observed at a concentration of 800μg/ml. MDAP-2 also had lower cytotoxicity to mammalian cells; IC50 values for HEK-293 cells, VERO cells, and IPEC-J2 cells were greater than 1000 μg/ml. The circular dichroism (CD) spectra showed that the peptide most- ly has α-helical properties and some β-fold structure in water and in membrane-like conditions. MDAP-2 is therefore a promising antibacterial agent against Gram-negative bacteria. To deter- mine the antibacterial mechanism(s) of action, fluorescent probes, flow cytometry, and transmis- sion electron microscopy (TEM) were used to study the effects of MDAP-2 on membrane perme- ability, polarization ability, and integrity of Gram-negative bacteria. The results indicated that the peptide caused membrane depolarization, increased membrane permeability, and destroyed membrane integrity. In conclusion, MDAP-2 is a broad-spectrum, lower hemolytic activity, and lower cytotoxicity antibacterial peptide, which is mainly effective on Gram-negative bacteria. It exerts its antimicrobial effects by causing bacterial cytoplasm membrane depolarization, increas- ing cell membrane permeability and disturbing the membrane integrity of Gram-negative bacte- ria. MDAP-2 may offer a new strategy to for defense against Gram-negative bacteria.

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Authors and Affiliations

Z. Pei
X. Ying
Y. Tang
L. Liu
H. Zhang
S. Liu
D. Zhang
K. Wang
L. Kong
Y. Gao
H. Ma
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Abstract

In this study, we developed a SYBR Green I real-time PCR method for the rapid and sensitive detection of novel porcine parvovirus 7 (PPV7). Specific primers were designed based on the highly conserved region within the Capsid gene of PPV7. The established method was 1,000 times more sensitive than the conventional PCR method and had a detection limit of 35.6 copies. This method was specific and had no cross-reactions with PCV2, PCV3, PRV, PEDV, PPV1, and PPV6. Experiments testing the intra and interassay precision demonstrated a high reproducibility. Testing the newly established method with 200 clinical samples revealed a detection rate up to 17.5% higher than that of the conventional PCR assay. The established method could provide technical support for clinical diagnosis and epidemiological investigation of PPV7.
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Authors and Affiliations

Y.D. Li
1
Z.D. Yu
2
C.X. Bai
2
D. Zhang
2
P. Sun
2
M.L Peng
2
H. Liu
3
ORCID: ORCID
J. Wang
4
Y. Wang
2
ORCID: ORCID

  1. Municipal Key Laboratory of Virology, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, PR China
  2. Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
  3. Anhui Animal Diseases Prevention and Control Center and Key Laboratory of Veterinary Pathobiology and Disease Prevention and Control of Anhui Province, Hefei 230091, PR China
  4. Animal Husbandry Base Teaching and Research Section, College of Animal Science and Technology, Hebei North University, Hebei 075000, PR China

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