Towards the elimination of hepatitis C: implementation of reflex testing in Andalusia.

BACKGROUND AND AIM: undiagnosed hepatitis C virus (HCV) infection and/or inadequate access to care are barriers to the elimination of HCV. Reflex testing has proven to facilitate referral to care, treatment and viral elimination. In this study, a reflex testing program was implemented in Andalusia and its impact on access to care was evaluated. PATIENTS AND METHODS: an observational, retrospective and prospective study was performed across diagnostic laboratories responsible for HCV diagnosis in southern Spain. After surveying the barriers to performing reflex testing, the number of patients that were not referred for care in 2016 was retrospectively studied (pre-reflex cohort). Subsequently, several measures were proposed to overcome the identified barriers. Finally, reflex testing was implemented and its impact evaluated. RESULTS: the pre-reflex cohort included information from 1,053 patients. Slightly more than half of the patients (n = 580; 55%) visited a specialist for treatment evaluation during a median period of 71 days (interquartile range = 35-134) since the date of diagnosis. The post-reflex cohort (September 2017 to March 2018) included 623 patients. Only 17% (n = 106) of the patients had not been referred for care or evaluated for treatment in a median period of 52 days (interquartile range = 28-86). CONCLUSIONS: in 2016, nearly half of new HCV diagnoses in southern Spain were not referred for care. Barriers to the implementation of reflex testing were overcome in our study. Moreover, this strategy was effectively implemented in 2017. Reflex testing contributed to improving referral for care. This program will contribute to the micro-elimination of hepatitis C in Spain.

[Laboratory diagnosis of HIV infection, viral tropism and resistance to antiretrovirals]. / Diagnóstico de laboratorio de la infección por el VIH, del tropismo viral y de las resistencias a los antirretrovirales.

The accurate diagnosis of HIV infection demands that to consider a positive result, at least three assays with different antigenic base should be used, one of them, Western-Blot being mandatory for confirmation. Fourth generation ELISAs shorten the window phase to 13-15 days, as they now include p24 antigen detection. Proviral DNA or Viral RNA detection by molecular methods have proved useful for addressing complex situations in which serology was inconclusive. Viral load (HIV-RNA) is routinely used to follow-up HIV infected patients and is used for treatment initiation decisions. It is also used to monitor viral failure. When this happens, resistance tests are needed to guide treatment changes. Resistance is also used to assess the transmission of drug resistance to newly diagnosed patients. Finally, before using an anti-CCR5 drug, viral tropism needs to be determined. This can be done using genotypic tests, widely available in many HIV labs, or phenotypic tests, only available at certain sites.

Breast Abscesses Caused by Anaerobic Microorganisms: Clinical and Microbiological Characteristics.

The objectives of this study were to report the antimicrobial susceptibility of 35 clinically significant anaerobic bacteria isolated from breast abscesses between March 2017 and February 2020 in a tertiary hospital in Granada (Spain) and to describe key clinical features of the patients. Species identification was performed mainly by MALDI-TOF MS. Antimicrobial susceptibility tests were carried out against benzylpenicillin, amoxicillin-clavulanic acid, imipenem, moxifloxacin, clindamycin, metronidazole, and piperacillin-tazobactam using the gradient diffusion technique and European Committee on Antimicrobial Susceptibility Testing EUCAST breakpoints (except for moxifloxacin). The most frequent anaerobes were Finegoldia magna (31.4%; n = 11), Actinomyces spp. (17.1%; n = 6), Propionibacterium spp. (17.1%; n = 6), and Prevotella spp. (14.2%; n = 5). Imipenem, amoxicillin-clavulanic acid, and piperacillin-tazobactam were universally active against all genera tested. High overall resistance rates to clindamycin were observed, especially for Gram-positive anaerobic cocci (56.2%) and Gram-positive anaerobic bacilli (38.4%). High resistance rates to metronidazole were also observed for Gram-positive (76.9%) and Gram-negative anaerobic bacilli (50%). High resistance rates to moxifloxacin were found for Gram-negative anaerobic bacilli (50%) and Gram-positive anaerobic cocci (31.2%). No breast abscess cases of Bacteroides spp. were detected. Routine antimicrobial susceptibility testing for anaerobes in breast abscesses may contribute to allow empirical therapies to be selected in accordance with local data on resistant strains.

Clinically relevant transmitted drug resistance to first line antiretroviral drugs and implications for recommendations.

BACKGROUND: The aim was to analyse trends in clinically relevant resistance to first-line antiretroviral drugs in Spain, applying the Stanford algorithm, and to compare these results with reported Transmitted Drug Resistance (TDR) defined by the 2009 update of the WHO SDRM list. METHODS: We analysed 2781 sequences from ARV naive patients of the CoRIS cohort (Spain) between 2007-2011. Using the Stanford algorithm "Low-level resistance", "Intermediate resistance" and "High-level resistance" categories were considered as "Resistant". RESULTS: 70% of the TDR found using the WHO list were relevant for first-line treatment according to the Stanford algorithm. A total of 188 patients showed clinically relevant resistance to first-line ARVs [6.8% (95%Confidence Interval: 5.8-7.7)], and 221 harbored TDR using the WHO list [7.9% (6.9-9.0)]. Differences were due to a lower prevalence in clinically relevant resistance for NRTIs [2.3% (1.8-2.9) vs. 3.6% (2.9-4.3) by the WHO list] and PIs [0.8% (0.4-1.1) vs. 1.7% (1.2-2.2)], while it was higher for NNRTIs [4.6% (3.8-5.3) vs. 3.7% (3.0-4.7)]. While TDR remained stable throughout the study period, clinically relevant resistance to first line drugs showed a significant trend to a decline (p = 0.02). CONCLUSIONS: Prevalence of clinically relevant resistance to first line ARVs in Spain is decreasing, and lower than the one expected looking at TDR using the WHO list. Resistance to first-line PIs falls below 1%, so the recommendation of screening for TDR in the protease gene should be questioned in our setting. Cost-effectiveness studies need to be carried out to inform evidence-based recommendations.

Improving Clinical Laboratory Efficiency: Introduction of Systems for the Diagnosis and Monitoring of HIV Infection.

Since the first tests for identifying individuals with suspected human immunodeficiency virus (HIV) infection were introduced in the mid-1980s, diagnostic virology testing has greatly evolved. The technological advances, automating in the laboratories and the advances in molecular biology techniques have helped introduce invaluable laboratory methods for managing HIV patients. Tests for diagnosis, specially for screening HIV antibodies, are now fully automated; in the same way, tests for monitoring HIV viral load (HIV RNA copies/ml of plasma), which is used for monitoring infection and response to antiretroviral treatment, are also fully automated; however, resistance testing, tropism determination and minor variant detection, which are used to make decisions for changing antiretroviral treatment regimens in patients failing therapy, still remain highly laborious and time consuming. This chapter will review the main aspects relating to the automating of the methods available for laboratory diagnosis as well as for monitoring of the HIV infection and determination of resistance to antiretrovirals and viral tropism.

Transmission of HIV drug resistance and non-B subtype distribution in the Spanish cohort of antiretroviral treatment naïve HIV-infected individuals (CoRIS).

CoRIS is an open multicentre cohort of HIV seroprevalent ARV-naïve subjects who began treatment at 32 Spanish healthcare centres from January 2004. Up to November 2008, a total of 683 FASTA format sequences, encoding the HIV protease and reverse transcriptase (RT) derived from plasma samples at entry into the cohort, had been obtained for examination of transmitted drug resistance (TDR) and HIV clade. TDR was found in 8.5% of the patients (4.4% NRTIs, 4% NNRTIs, 2.2% PIs). The most prevalent resistance mutations were: T215 revertants (3.8%), D67NG (1.3%), K219QENR (1.2%) and M41L (1%), for NRTIs; K103N (3.2%), for NNRTIs; I54VLMSAT, M46I and L90M (0.7%), for PIs. Non-B subtypes were recognized in 104 patients (15.2%) and were more common in Sub-Saharan Africans (15/17, 88.2%), Eastern Europeans (7/12, 58.3%) and Northern Africans (8/16, 50%) than among Spaniards (53/479, 11%) (p<0.001). The most prevalent non-B subtype was CRF02_AG (4.4%), followed by subtype D (1.9%), CRF03_AB (1.5%), CRF07_BC and subtype F1 (1%). A trend was observed for the transmission of non-B subtypes to increase and for TDR to decrease.

Predicting antiretroviral drug resistance from the latest or the cumulative genotype.

BACKGROUND: This study evaluates the added benefit when estimating antiretroviral drug resistance of combining all available resistance test results in a cumulative genotype relative to using the latest genotype alone. METHODS: The prevalence of resistance and genotypic sensitivity scores (GSS) predicted by the latest and the cumulative genotype, together with virological outcomes after the latest genotype, were measured in treatment-experienced patients who underwent ≥2 resistance tests in 1999-2008. RESULTS: Comparing the latest with the cumulative genotype in 227 patients, 4 (1.7%) versus 0 (0.0%) showed no major resistance mutations, whereas 74 (32.6%) versus 46 (20.3%), 88 (38.8%) versus 76 (33.5%) and 61 (26.9%) versus 105 (46.3%) showed single-class, dual-class and triple-class resistance mutations, respectively. The median (IQR) number of fully or partially active drugs was 6 (5-6) versus 5 (4-6) for the nucleoside/nucleotide reverse transcriptase inhibitors, 3 (1-3) versus 1 (1-3) for the non-nucleoside reverse transcriptase inhibitors and 7 (7-7) versus 7 (7-7) for the protease inhibitors, respectively. Among 163 patients who started a new regimen after the latest genotype, both the latest and the cumulative GSS were predictive of early (≤24 weeks) virological responses. The GSS decreased by median 1 unit (IQR 0.5-1.0) in the cumulative genotype and larger differences relative to the latest genotype corresponded to smaller decreases in viral load. CONCLUSIONS: The cumulative genotype offers a more comprehensive evaluation of the burden of resistance. This approach can guide small but appreciable improvements in the selection of antiretroviral regimens for treatment-experienced patients.

Towards the elimination of hepatitis C: implementation of reflex testing in Andalusia

BACKGROUND AND AIM: undiagnosed hepatitis C virus (HCV) infection and/or inadequate access to care are barriers to the elimination of HCV. Reflex testing has proven to facilitate referral to care, treatment and viral elimination. In this study, a reflex testing program was implemented in Andalusia and its impact on access to care was evaluated. PATIENTS AND METHODS: an observational, retrospective and prospective study was performed across diagnostic laboratories responsible for HCV diagnosis in southern Spain. After surveying the barriers to performing reflex testing, the number of patients that were not referred for care in 2016 was retrospectively studied (pre-reflex cohort). Subsequently, several measures were proposed to overcome the identified barriers. Finally, reflex testing was implemented and its impact evaluated. RESULTS: the pre-reflex cohort included information from 1,053 patients. Slightly more than half of the patients (n = 580; 55%) visited a specialist for treatment evaluation during a median period of 71 days (interquartile range = 35-134) since the date of diagnosis. The post-reflex cohort (September 2017 to March 2018) included 623 patients. Only 17% (n = 106) of the patients had not been referred for care or evaluated for treatment in a median period of 52 days (interquartile range = 28-86). CONCLUSIONS: in 2016, nearly half of new HCV diagnoses in southern Spain were not referred for care. Barriers to the implementation of reflex testing were overcome in our study. Moreover, this strategy was effectively implemented in 2017. Reflex testing contributed to improving referral for care. This program will contribute to the micro-elimination of hepatitis C in Spain

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Diagnóstico de laboratorio de la infección por el VIH, del tropismo viral y de las resistencias a los antirretrovirales / Laboratory diagnosis of HIV infection, viral tropism and resistance to

En el diagnóstico de la infección VIH, para considerar un resultado positivo, se recomienda el uso de tres técnicas con distinto principio o base antigénica, siendo obligado que para la confirmación una de ellas sea el Western Blot. Las técnicas serológicas de cuarta generación acortan el período ventana a 13-15 días, debido a que incluyen la detección de antígeno-p24. La detección del genoma-VIH (ADN proviral/ARN) complementa al diagnóstico serológico en situaciones complejas. La viremia plasmática (carga viral) se utiliza para el seguimiento de los pacientes infectados por VIH, para contribuir a la decisión del inicio de tratamiento y para comprobar el fallo virológico al régimen antirretroviral en uso. Las pruebas de resistencia se utilizan para guiar el cambio de tratamiento, y para detectar la transmisión de cepas resistentes en los nuevos diagnósticos. Antes de utilizar un antagonista de CCR5 hay que determinar el tropismo viral; para ello, se pueden utilizar métodos genotípicos, accesibles a los laboratorios de diagnóstico, o fenotípicos, de más difícil acceso (AU)

The accurate diagnosis of HIV infection demands that to consider a positive result, at least three assays with different antigenic base should be used, one of them, Western-Blot being mandatory for confirmation. Fourth generation ELISAs shorten the window phase to 13-15 days, as they now include p24 antigen detection. Proviral DNA or Viral RNA detection by molecular methods have proved useful for addressing complex situations in which serology was inconclusive. Viral load (HIV-RNA) is routinely used to follow up HIV infected patients and is used for treatment initiation decisions. It is also used to monitor viral failure. When this happens, resistance tests are needed to guide treatment changes. Resistance is also used to assess the transmission of drug resistance to newly diagnosed patients. Finally, before using ananti-CCR5 drug, viral tropism needs to be determined. This can be done using genotypic tests, widely available in many HIV labs, or phenotypic tests, only available at certain sites (AU)