Diagnosis of porcine circovirus type 2 infection with a combination of immunomagnetic beads, single-domain antibody, and fluorescent quantum dot probes

Diagnosis of porcine circovirus type 2 infection with a combination of immunomagnetic beads,... The use of a specific antibody conjugated with nanobeads, forming immunomagnetic nanobeads (IMNBs), has been demonstrated to be useful for the capture and detection of viruses. In this study, IMNBs functionalized with a single-domain antibody against the capsid protein (Cap) of porcine circovirus type 2 (PCV2), hereafter denoted as psdAb, were evaluated and used to capture PCV2. Quantum dots (QDs) conjugated with psdAb were used as a fluorescence probe to visualize PCV2 captured by IMNBs. The specificity and sensitivity of this method were further evaluated using common pathogens of pig viral disease and PCV2. To assess its practicality, clinical samples were tested in this study. The results showed that 2.57 ± 0.13 mg Cap or 0.97 ± 0.064 × 106 copies of PCV2 particles could be captured by 1 mg of IMNBs in 30 min. This suggests that the IMNBs have the ability to efficiently capture PCV2 with good specificity, as there was no cross-reaction with other pathogens, and with strong sensitivity, with a detection limit as low as 103 copies/ml of PCV2 particles. Moreover, PCV2 in inguinal lymph node, lung, spleen, serum, and fecal samples was successfully detected by IMNBs. The results demonstrate that this method is promising for the rapid and effective detection of PCV2 in complex clinical samples. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Diagnosis of porcine circovirus type 2 infection with a combination of immunomagnetic beads, single-domain antibody, and fluorescent quantum dot probes

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Publisher
Springer Journals
Copyright
Copyright © 2015 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-015-2508-x
Publisher site
See Article on Publisher Site

Abstract

The use of a specific antibody conjugated with nanobeads, forming immunomagnetic nanobeads (IMNBs), has been demonstrated to be useful for the capture and detection of viruses. In this study, IMNBs functionalized with a single-domain antibody against the capsid protein (Cap) of porcine circovirus type 2 (PCV2), hereafter denoted as psdAb, were evaluated and used to capture PCV2. Quantum dots (QDs) conjugated with psdAb were used as a fluorescence probe to visualize PCV2 captured by IMNBs. The specificity and sensitivity of this method were further evaluated using common pathogens of pig viral disease and PCV2. To assess its practicality, clinical samples were tested in this study. The results showed that 2.57 ± 0.13 mg Cap or 0.97 ± 0.064 × 106 copies of PCV2 particles could be captured by 1 mg of IMNBs in 30 min. This suggests that the IMNBs have the ability to efficiently capture PCV2 with good specificity, as there was no cross-reaction with other pathogens, and with strong sensitivity, with a detection limit as low as 103 copies/ml of PCV2 particles. Moreover, PCV2 in inguinal lymph node, lung, spleen, serum, and fecal samples was successfully detected by IMNBs. The results demonstrate that this method is promising for the rapid and effective detection of PCV2 in complex clinical samples.

Journal

Archives of VirologySpringer Journals

Published: Jul 9, 2015

References

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