Journal Information
Vol. 15. Issue 6.
Pages 547-552 (November - December 2011)
Share
Share
Download PDF
More article options
Vol. 15. Issue 6.
Pages 547-552 (November - December 2011)
Original article
Open Access
Magnetic bead technology for viral RNA extraction from serum in blood bank screening
Visits
3534
Guilherme Ambrozio Albertoni1,
Corresponding author
albertonig@nefro.epm.br

Correspondence to: Av. Jandira, 1260 Indianópolis 04080-006 São Paulo, SP Brazil Phone: +55 11 5055-6588 R-39 Fax: +55 11 5055-6588.
, Carine Prisco Arnoni1, Patricia Regina Barboza Araujo1, Sheila Siqueira Andrade1, Fabrício Oliveira Carvalho2, Manoel João Batista Castello Girão3, Nestor Schor4, José Augusto Barreto5
1 Colsan – Associação Beneficente de Coleta de Sangue, São Paulo, SP, Brazil
2 Colsan – Associação Beneficente de Coleta de Sangue, São Paulo, SP, Brazil
3 Colsan – Associação Beneficente de Coleta de Sangue, São Paulo, SP, Brazil
4 Universidade Federal de São Paulo (UNIFESP), SP, Brazil
5 Head, Colsan – Associação Beneficente de Coleta de Sangue, São Paulo, SP, Brazil
This item has received

Under a Creative Commons license
Article information
Abstract

Nucleic acid amplification testing (NAT) was recently recommended by Brazilian legislation and has been implemented at some blood banks in the city of São Paulo, Brazil, in an attempt to reduce blood-born transmission of human immunodeficiency virus (HIV) and hepatitis C virus.

Objective

Manual magnetic particle-based extraction methods for HIV and HCV viral nucleic acids were evaluated in combination with detection by reverse transcriptase -polymerase chain reaction (RT-PCR) one-step.

Methods

Blood donor samples were collected from January 2010 to September 2010, and minipools of them were submitted to testing. ELISA was used for the analysis of anti-HCV/HIV antibodies. Detection and amplification of viral RNA was performed using real-time PCR.

Results

Out of 20.808 samples screened, 53 samples (29 for HCV and 24 for HIV) were confirmed as positive by serological and NAT methods.

Conclusion

The manual magnetic bead-based extraction in combination with real-time PCR detection can be used to routinely screen blood donation for viremic donors to further increase the safety of blood products.

Keywords:
HCV
HIV
infection
reverse transcriptase polymerase chain reaction
blood banks
Full text is only aviable in PDF
References
[1.]
H.J. Alter, M. Houghton.
Hepatitis C virus and eliminating post transfusion hepatitis.
Nat Med, 6 (2000), pp. 1082-1086
[2.]
S.L. Stramer, S.A. Glynn, S.H. Kleinman, et al.
Detection of HIV-1 and HCV infections among antibody-negative blood donors by nucleic acid-amplification testing.
N Engl J Med, 351 (2004), pp. 760-768
[3.]
W.K. Roth, M. Weber, E. Seifried.
Feasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting.
Lancet, 53 (1999), pp. 359-363
[4.]
S. Laperche.
Blood safety and nucleic acid testing in Europe.
Euro Survey, 10 (2005), pp. 3-4
[5.]
J. Coste, H.W. Reesink, C.P. Engelfriet, S. Laperche.
Implementation of donor screening for infectious agents transmitted by blood by nucleic acid technology.
[6.]
C. Velati, L. Fomiatti, L. Baruffi, L. Romano, A. Zanetti.
Impact of nucleic acid amplification technology (NAT) in Italy in the three years folowing implementation (2001–2003).
Euro Surveill, 10 (2005), pp. 12-14
[7.]
B.R. Jackson, M.O. Busch, S.L. Stramer, J.P. AuBuchon.
The costeffectiveness of NAT for HIV, HCV and HBV in whole-blood donations.
Transfusion, 43 (2003), pp. 721-729
[8.]
W.K. Roth, S. Buhr, C. Drosten, E. Seifried.
NAT and viral safety in blood transfusion.
Vox Sang, 78 (2000), pp. 257-259
[9.]
S. Braham, J. Gandhi, S. Beard, B. Cohen.
Evaluation of the Roche lightCycler parvovirus B19 quantification kit for the diagnosis of parvovirus B19 infections.
J Clin Virol, 31 (2004), pp. 5-10
[10.]
J.P. Allain.
Transfusion risks of yesterday and of today.
Transfus Clin Biol, 10 (2003), pp. 1-5
[11.]
E.C. Sabino, N.A. Salles, A. Sáez-Alquezar, G.R. Santos, D.F. Chamone, M.P. Busch.
Estimated risk of transfusion-transmitted HIV infection in São Paulo.
Brazil. Transfusion, 39 (1999), pp. 1152-1153
[12.]
S. Wendel, R.M. Fachini, J.E. Levi, et al.
A single window-period donation detected by human immunodeficiency vírus p24 antigen after 5 years of routine screening in a group of Brazilian blood banks.
Vox Sang, 83 (2002), pp. 309-312
[13.]
C.C. Barreto, E.C. Sabino, T.T. Gonçalvez, et al.
Prevalence, incidence and residual risk of human immunodeficiency virus among community and replacement first-time blood donors in São Paulo, Brazil.
Transfusion, 45 (2005), pp. 1709-1714
[14.]
Ministério da Saúde, Secretaria de Atenção à Saúde, Departamento de Atenção Especializada, Coordenação Geral de Sangue e Hemoderivados. Estágio Atual da Implantação dos Testes de Ácido Nucléico para HIV e HCV – NAT HIV/HCV na rede de Serviços de Hemoterapia do Brasil. Nota técnica, 2010.
[15.]
M.K. Hourfar, M. Schimidt, E. Seifried, W.K. Roth.
Evaluation of an automated high-volume extraction method for viral nucleic acids in comparison to a manual procedure with preceding enrichment.
[16.]
J.L. Fleiss.
Statistical methods for rates and proportion.
2nd ed., John Wiley & Sons, (1981),
[17.]
J.R. Landis, G.G. Koch.
The measurement of observer agreement for categorical data.
Biometrics, 33 (1977), pp. 159-174
[18.]
J.L. Gerberding, D.W. Fleming, D.E. Snider.
Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus.
Epidemiology Program Office, Centers for Disease control and Prevention (CDC), Department of Health and Human Services, 52 (2003), pp. 1-15
[19.]
R. Boom, J.C.R. Sol, M.M. Salimans, C.L. Jansen, P.M. Wertheim-van Dillen, J. Van Der Noordaa.
Rapid and simple method for purification of nucleic acids.
J Clin Microbiol, 28 (1990), pp. 495-503
[20.]
B. Vogelstein, D. Gillespie.
Preparative and purification of DNA from agarose.
Proc Natl Acad Sci USA, 76 (1979), pp. 615-669
[21.]
F.J. Castellino, R. Barker.
The denaturing effectiveness of guanidinium, carbamoylguanidinium, and guanylguanidinium salts.
Biochemistry, 7 (1968), pp. 4135-4138
[22.]
J.M. Chirgwin, A.E. Przbyla, R.J. MacDonald, W.J. Rutter.
Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
Biochemistry, 8 (1979), pp. 5294-5299
[23.]
K.A. Melzak, C.S. Sherwood, R.F.B. Turner, C.A. Haynes.
Driving forces for DNA adsorption to silica in perchlorate solutions.
J Colloid Interface Sci, 181 (1996), pp. 635-644
[24.]
L. Pichl, A. Heitmann, P. Herzog, J. Oster, H. Smets, V. Schottstedt.
Magnetic bead technology in viral RNA and DNA extraction from plasma minipools.
Transfusion, 45 (2005), pp. 1106-1110
[25.]
M.K. Hourfar, U. Michelsen, M. Schmidt, A. Berger, E. Seifried, W.K. Roth.
High-throughput purification of viral RNA based on novel aqueous chemistry for nucleic acid isolation.
Clin Chemistry, 7 (2005), pp. 1217-1222
[26.]
L. Jarvis, J. Becker, A. Tender, et al.
Evaluation of the Roche cobas 201 system and cobas TaqScreen multiplex test for blood screening: European multicenter study.
Transfusion, 48 (2008), pp. 1853-1861
[27.]
G.A. Albertoni, C.P. Arnoni, P.R.B. Araújo, F.O. Carvalho, J.A. Barreto.
Signal to-cut-off (S/CO) ratio and detection of HCV genotype I by real time PCR one-step method: is there any direct relationship?.
Braz J Infect Dis, 14 (2010), pp. 147-152
Copyright © 2011. Elsevier Editora Ltda.. All rights reserved
Download PDF
The Brazilian Journal of Infectious Diseases
Article options
Tools