Journal article
A novel, rapid method to detect infectious HIV-1 from plasma of persons infected with HIV-1
A Cornall, L Sharma, A Solomon, PR Gorry, SM Crowe, PU Cameron, SR Lewin
Journal of Virological Methods | ELSEVIER | Published : 2010
Abstract
Efficient isolation of replication-competent virus from plasma of patients infected with HIV-1 is needed to characterize important clinical parameters of virus. However, addition of plasma to in vitro cultures results in clot formation. Blood from HIV-1 infected patients was collected in the presence of three commonly used anticoagulants (ACD, heparin and EDTA) and plasma was isolated. Plasma was then used to infect HIV-1 indicator cell lines (TZM-bl and GHOST) with spinoculation in the presence or absence of additional heparin and positively charged polymers. The presence of additional heparin during inoculation significantly reduced clot formation without affecting the sensitivity of HIV-1..
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Funding Acknowledgements
This work was supported by a grant from the Immunovirology Research Network (IVRN), Australian Centre for HIV and Hepatitis, Population Health Division, Commonwealth Government, Australia. PRG is the recipient of an Australian National Health and Medical Research Council Level 2 Biomedical Career Development Award. SRL is an NHMRC Practitioner Fellow. SMC is an NHMRC Principal Research Fellow. The authors are grateful to Candida da Fonseca Pereira of the Macfarlane Burnet Institute for her assistance with the use of the confocal microscope, Geza Paukovics of the Macfarlane Burnet Institute and Monash University for his assistance with flow cytometry and Clinical Research Nurses, Infectious Diseases Unit, Alfred Hospital for the recruitment of the patients for the study.