Avaliação da carga viral do vírus da imunodeficiência humana (HIV) em plasma pobre e rico em plaquetas
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Data
2021-05-28
Autores
Schnoor, Suzana Kague
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Universidade Estadual Paulista (Unesp)
Resumo
A Aids é consequência da infecção causada pelo vírus da imunodeficiência humana (HIV). Os principais alvos do vírus são linfócitos T CD4+ e os macrófagos, porém já foi demonstrado que as plaquetas interagem com o vírus mesmo não expressando a molécula CD4 em sua superfície celular. Estudos recentes demonstraram que as plaquetas não só interagem com o HIV como são reservatórios de vírus infectantes, capazes de transmiti-lo a outras células suscetíveis. A determinação da carga viral do HIV é considerada padrão-ouro para avaliação da eficácia do tratamento antirretroviral e detecção precoce de problemas de adesão em pessoas vivendo com HIV, sendo realizada utilizando a técnica de transcrição reversa seguida da Reação em Cadeia da Polimerase em tempo real (RT-qPCR) em amostras de plasma pobre em plaquetas, de acordo com o protocolo estabelecido pelo Ministério da Saúde. O objetivo do presente trabalho foi avaliar comparativamente a carga viral do HIV no plasma pobre e rico em plaquetas em amostras de sangue de trinta pacientes, sem tratamento antirretroviral, portadores do vírus e que foram atendidos pelo Laboratório de Biologia Molecular do Hospital das Clínicas da Faculdade de Medicina de Botucatu. Foram utilizados dois protocolos de centrifugação para obtenção do plasma pobre e rico em plaquetas, seguidos de quantificação da carga viral através da metodologia Abbott RealTime HIV-1 (Abbott Molecular, Illinois, USA). Os resultados demonstraram não haver diferença estatística significativa entre os dois grupos avaliados (plasma rico e pobre em plaquetas). Esperava-se que o plasma rico em plaquetas apresentasse uma carga viral maior do que o plasma pobre, uma vez que as mesmas abrigam o vírus, porém essa diferença não foi observada. No entanto, a metodologia pode explicar estes resultados. Os resultados sugerem testar novos protocolos, como utilização de colunas de sílica para extração do RNA viral, uma vez que o protocolo atual utiliza partículas magnéticas que não são especificas para ácidos nucleicos e podem sofrer interferências de outras moléculas presentes na amostra, e outros protocolos de centrifugação para obtenção de plasma rico em plaquetas como descritos em outros estudos.
AIDS is a consequence of infection caused by the human immunodeficiency virus (HIV). The main target of the virus is the immune system, especially CD4 + T lymphocytes and macrophages, although platelets have been shown to interact with the virus even though not expressing the CD4 molecule on their cell surface. Recent studies have shown that platelets not only interact with HIV but are also reservoirs of infecting viruses, capable of transmitting it to other susceptible cells. Laboratory monitoring of HIV viral load is considered the gold standard to assess the effectiveness of antiretroviral treatment and early detection of adherence problems in people living with HIV, and the quantification of viral load is performed using the reverse transcription technique followed by Reaction in Polymerase Chain in real time (RT-qPCR) in platelet-poor plasma samples, according to the protocol established by the Ministry of Health. Two centrifuge protocols were used to obtain poor and platelet-rich plasma, followed by quantification of viral load using Abbott RealTime HIV-1 (Abbott Molecular, Illinois, USA) methodology. The objective of the present study was to evaluate comparatively the HIV viral load in the poor and platelet-rich plasma in blood samples from patients, without antiretroviral treatment, carrying the virus and who were attended by the Molecular Biology Laboratory of the Hospital das Clínicas, Botucatu School of Medicine. The results have shown that there is no statistically significant difference between the two groups evaluated. Platelet-rich plasma was expected to have a higher viral load than poor plasma, once they fight the virus, but this difference was not observed. Therefore, we have suggest testing new protocols, such as the use of silica columns to extract viral RNA, once the current protocol uses magnetic particles that are not specific for nucleic acids and may suffer interference from other molecules present in the sample, and other centrifugation protocols to get platelet-rich plasma as described in other studies.
AIDS is a consequence of infection caused by the human immunodeficiency virus (HIV). The main target of the virus is the immune system, especially CD4 + T lymphocytes and macrophages, although platelets have been shown to interact with the virus even though not expressing the CD4 molecule on their cell surface. Recent studies have shown that platelets not only interact with HIV but are also reservoirs of infecting viruses, capable of transmitting it to other susceptible cells. Laboratory monitoring of HIV viral load is considered the gold standard to assess the effectiveness of antiretroviral treatment and early detection of adherence problems in people living with HIV, and the quantification of viral load is performed using the reverse transcription technique followed by Reaction in Polymerase Chain in real time (RT-qPCR) in platelet-poor plasma samples, according to the protocol established by the Ministry of Health. Two centrifuge protocols were used to obtain poor and platelet-rich plasma, followed by quantification of viral load using Abbott RealTime HIV-1 (Abbott Molecular, Illinois, USA) methodology. The objective of the present study was to evaluate comparatively the HIV viral load in the poor and platelet-rich plasma in blood samples from patients, without antiretroviral treatment, carrying the virus and who were attended by the Molecular Biology Laboratory of the Hospital das Clínicas, Botucatu School of Medicine. The results have shown that there is no statistically significant difference between the two groups evaluated. Platelet-rich plasma was expected to have a higher viral load than poor plasma, once they fight the virus, but this difference was not observed. Therefore, we have suggest testing new protocols, such as the use of silica columns to extract viral RNA, once the current protocol uses magnetic particles that are not specific for nucleic acids and may suffer interference from other molecules present in the sample, and other centrifugation protocols to get platelet-rich plasma as described in other studies.
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Palavras-chave
HIV, Carga viral, Plasma rico em plaquetas, Plasma pobre em plaquetas, Plaquetas, Viral load, Platelet-rich plasma, Platelet-poor plasma, Platelets