BACKGROUND: Numerous studies investigating clinical significance of HIV-1 minimal residual viremia (MRV) suggest potential utility of assays more sensitive than those routinely used to monitor viral suppression. However currently available methods, based on different technologies, show great variation in detection limit and input plasma volume, and generally suffer from lack of standardization. OBJECTIVES: In order to establish new tools suitable for routine quantification of minimal residual viremia in patients under virological suppression, some modifications were introduced into standard procedure of the Abbott RealTime HIV-1 assay leading to a "modified" and an "ultrasensitive" protocols. STUDY DESIGN: The following modifications were introduced: calibration curve extended towards low HIV-1 RNA concentration; 4 fold increased sample volume by concentrating starting material; reduced volume of internal control; adoption of "open-mode" software for quantification. Analytical performances were evaluated using the HIV-1 RNA Working Reagent 1 for NAT assays (NIBSC). Both tests were applied to clinical samples from virologically suppressed patients. RESULTS: The "modified" and the "ultrasensitive" configurations of the assay reached a limit of detection of 18.8 (95% CI: 11.1-51.0 cp/mL) and 4.8 cp/mL (95% CI: 2.6-9.1 cp/mL), respectively, with high precision and accuracy. In clinical samples from virologically suppressed patients, "modified" and "ultrasensitive" protocols allowed to detect and quantify HIV RNA in 12.7% and 46.6%, respectively, of samples resulted "not-detectable", and in 70.0% and 69.5%, respectively, of samples "detected <40 cp/mL" in the standard assay. CONCLUSIONS: The "modified" and "ultrasensitive" assays are precise and accurate, and easily adoptable in routine diagnostic laboratories for measuring MRV.
BACKGROUND: Numerous studies investigating clinical significance of HIV-1 minimal residual viremia (MRV) suggest potential utility of assays more sensitive than those routinely used to monitor viral suppression. However currently available methods, based on different technologies, show great variation in detection limit and input plasma volume, and generally suffer from lack of standardization. OBJECTIVES: In order to establish new tools suitable for routine quantification of minimal residual viremia in patients under virological suppression, some modifications were introduced into standard procedure of the Abbott RealTime HIV-1 assay leading to a "modified" and an "ultrasensitive" protocols. STUDY DESIGN: The following modifications were introduced: calibration curve extended towards low HIV-1 RNA concentration; 4 fold increased sample volume by concentrating starting material; reduced volume of internal control; adoption of "open-mode" software for quantification. Analytical performances were evaluated using the HIV-1 RNA Working Reagent 1 for NAT assays (NIBSC). Both tests were applied to clinical samples from virologically suppressed patients. RESULTS: The "modified" and the "ultrasensitive" configurations of the assay reached a limit of detection of 18.8 (95% CI: 11.1-51.0 cp/mL) and 4.8 cp/mL (95% CI: 2.6-9.1 cp/mL), respectively, with high precision and accuracy. In clinical samples from virologically suppressed patients, "modified" and "ultrasensitive" protocols allowed to detect and quantify HIV RNA in 12.7% and 46.6%, respectively, of samples resulted "not-detectable", and in 70.0% and 69.5%, respectively, of samples "detected <40 cp/mL" in the standard assay. CONCLUSIONS: The "modified" and "ultrasensitive" assays are precise and accurate, and easily adoptable in routine diagnostic laboratories for measuring MRV.
Authors: Marta Álvarez Estévez; Natalia Chueca Porcuna; Vicente Guillot Suay; Alejandro Peña Monge; Fernando García García; Leopoldo Muñoz Medina; David Vinuesa García; Jorge Parra Ruiz; Jose Hernández-Quero; Federico García García Journal: J Clin Microbiol Date: 2013-02-06 Impact factor: 5.948
Authors: Luke C Swenson; Bryan Cobb; Anna Maria Geretti; P Richard Harrigan; Mario Poljak; Carole Seguin-Devaux; Chris Verhofstede; Marc Wirden; Alessandra Amendola; Jurg Boni; Thomas Bourlet; Jon B Huder; Jean-Claude Karasi; Snjezana Zidovec Lepej; Maja M Lunar; Odette Mukabayire; Rob Schuurman; Janez Tomazic; Kristel Van Laethem; Linos Vandekerckhove; Annemarie M J Wensing Journal: J Clin Microbiol Date: 2013-12-04 Impact factor: 5.948
Authors: Concepcion Casado; Cristina Galvez; Maria Pernas; Laura Tarancon-Diez; Carmen Rodriguez; Víctor Sanchez-Merino; Mar Vera; Isabel Olivares; Rebeca De Pablo-Bernal; Alberto Merino-Mansilla; Jorge Del Romero; Ramon Lorenzo-Redondo; Ezequiel Ruiz-Mateos; María Salgado; Javier Martinez-Picado; Cecilio Lopez-Galindez Journal: Sci Rep Date: 2020-02-05 Impact factor: 4.379
Authors: Oscar Blanch-Lombarte; Cristina Gálvez; Boris Revollo; Esther Jiménez-Moyano; Josep M Llibre; José Luís Manzano; Aram Boada; Judith Dalmau; Daniel E Speiser; Bonaventura Clotet; Julia G Prado; Javier Martinez-Picado Journal: J Clin Med Date: 2019-12-01 Impact factor: 4.241