Viruses 2014, 6(3), 1037-1048; doi:10.3390/v6031037
Article

Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus

1,†email, 2,†email, 3,†email, 2email, 3email, 1email, 4,5email, 3email, 3,5email and 3,5,* email
Received: 9 September 2013; in revised form: 14 January 2014 / Accepted: 17 January 2014 / Published: 4 March 2014
(This article belongs to the Section Animal Viruses)
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Abstract: The accuracy and sensitivity of PCR-based methods for detection of hepatitis A virus (HAV) are dependent on the methods used to separate and concentrate the HAV from the infected cells. The pH and ionic strength affect the binding affinity of the virus to cells. In this study, we initially investigated the effects of pH (4.0–10.0) and metal ions (Fe2+, Co2+, Cu2+, Mg2+, K+, and Ca2+) on the binding of HAV to oyster digestive cells. The lowest relative binding (RB) of HAV to the cells was found at pH 4.0 and in FeSO4 solution (64.6% and 68.1%, respectively). To develop an alternative to antibody-dependent immunomagnetic separation prior to detection of HAV using RT-PCR, the binding of HAV to five lectins, peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Ulex europaeus agglutinin (UEA-1) and soybean agglutinin (SBA), was evaluated using ELISAs. SBA showed significantly higher RB to HAV than the other lectins tested. In addition, HAV could be concentrated within 30 min using SBA-linked magnetic bead separation (SMS) prior to the RT-PCR assay. Our findings demonstrate the feasibility of using SMS combined with RT-PCR to detect HAV at dilutions ranging from 10−1–10−4 of a HAV stock (titer: 104 TCID50/mL).
Keywords: hepatitis A virus; soybean agglutinin linked-magnetic bead separation; lectin; oyster
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Ko, S.-M.; Kwon, J.; Vaidya, B.; Choi, J.S.; Lee, H.-M.; Oh, M.-J.; Bae, H.-J.; Cho, S.-Y.; Oh, K.-S.; Kim, D. Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus. Viruses 2014, 6, 1037-1048.

AMA Style

Ko S-M, Kwon J, Vaidya B, Choi JS, Lee H-M, Oh M-J, Bae H-J, Cho S-Y, Oh K-S, Kim D. Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus. Viruses. 2014; 6(3):1037-1048.

Chicago/Turabian Style

Ko, Sang-Mu; Kwon, Joseph; Vaidya, Bipin; Choi, Jong S.; Lee, Hee-Min; Oh, Myung-Joo; Bae, Hyeun-Jong; Cho, Se-Young; Oh, Kyung-Seo; Kim, Duwoon. 2014. "Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus." Viruses 6, no. 3: 1037-1048.


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