Repurposing Positive SARS-CoV-2 Antigen Test Devices for Variant Tracking.

From the very beginning of the SARS-CoV-2 pandemic, one of the very few common opinions was that to control the expansion of the virus as many as the possible test had to be done. Antigen tests, being affordable and easy and fast to use, represented a great opportunity to expand the testing capacities of many healthcare systems. However, in 2021 with the appearance of the new SARS-CoV-2 variants, variant tracking strategies had to be implemented, which often included needing a second test to determine the variant of the patients diagnosed with antigen tests or not taking these samples into consideration at all. Therefore, we proposed recovering the positive antigen test devices to include them in our routine variant tracking strategy. The recovered positive antigen test devices obtained from 1st April 2021 to 15the January 2022 were analysed following the variant tracking protocol in force. The results obtained were compared to the positive samples detected by RT-PCR which were processed for variant tracking during the same period. 21,304 samples were processed, 6297 from the recovered positive antigen devices and 15,007 from the standard nasopharyngeal swabs. Only 773 (3.63%) samples were no conclusive, 104 (1.65%) from the recovered antigen devices and 669 (4.46%) from the RT-PCR positive group. This difference was statistically significant (p < 0.01). Taking this into account the proposed method is suitable and very recommendable, as it is an important measure to have a better and immediate picture of the circulating variants in every community.

Identification of CRF66_BF, a New HIV-1 Circulating Recombinant Form of South American Origin.

Circulating recombinant forms (CRFs) are important components of the HIV-1 pandemic. Among 110 reported in the literature, 17 are BF1 intersubtype recombinant, most of which are of South American origin. Among these, all 5 identified in the Southern Cone and neighboring countries, except Brazil, derive from a common recombinant ancestor related to CRF12_BF, which circulates widely in Argentina, as deduced from coincident breakpoints and clustering in phylogenetic trees. In a HIV-1 molecular epidemiological study in Spain, we identified a phylogenetic cluster of 20 samples from 3 separate regions which were of F1 subsubtype, related to the Brazilian strain, in protease-reverse transcriptase (Pr-RT) and of subtype B in integrase. Remarkably, 14 individuals from this cluster (designated BF9) were Paraguayans and only 4 were native Spaniards. HIV-1 transmission was predominantly heterosexual, except for a subcluster of 6 individuals, 5 of which were men who have sex with men. Ten additional database sequences, from Argentina (n = 4), Spain (n = 3), Paraguay (n = 1), Brazil (n = 1), and Italy (n = 1), branched within the BF9 cluster. To determine whether it represents a new CRF, near full-length genome (NFLG) sequences were obtained for 6 viruses from 3 Spanish regions. Bootscan analyses showed a coincident BF1 recombinant structure, with 5 breakpoints, located in p17 gag , integrase, gp120, gp41-rev overlap, and nef, which was identical to that of two BF1 recombinant viruses from Paraguay previously sequenced in NFLGs. Interestingly, none of the breakpoints coincided with those of CRF12_BF. In a maximum likelihood phylogenetic tree, all 8 NFLG sequences grouped in a strongly supported clade segregating from previously identified CRFs and from the CRF12_BF "family" clade. These results allow us to identify a new HIV-1 CRF, designated CRF66_BF. Through a Bayesian coalescent analysis, the most recent common ancestor of CRF66_BF was estimated around 1984 in South America, either in Paraguay or Argentina. Among Pr-RT sequences obtained by us from HIV-1-infected Paraguayans living in Spain, 14 (20.9%) of 67 were of CRF66_BF, suggesting that CRF66_BF may be one of the major HIV-1 genetic forms circulating in Paraguay. CRF66_BF is the first reported non-Brazilian South American HIV-1 CRF_BF unrelated to CRF12_BF.

Epidemiological surveillance study of gonococcal infection in Northern Spain / Estudio de vigilancia epidemiológica de la infección gonocócica en el Norte de España

BACKGROUND: Treatment of gonorrhoea is threatened by antimicrobial resistance, and decreased susceptibility to recommended therapies is emerging. Thus, gonococcal infection (GI) is becoming a public health problem. The objectives of the present study were to monitor the antimicrobial sensitivity in Neisseria gonorrhoeae (NG) during 2011-2015 and to study their genogroups. METHODS: Antimicrobial susceptibility was studied by disc diffusion, in addition to the agar dilution method for cefixime and ceftriaxone and the Etest(R) for azithromycin. Genotyping was performed by the NG multi-antigen sequence typing (NG-MAST) method. Genogroups of closely related sequence types (STs) were defined. RESULTS: All the strains were susceptible to cefixime, ceftriaxone and gentamicin and 1.8% of the strains were resistant to azithromycin. A total of 531 STs and 6 genotypes (Gs) were identified during 2012-2015 period. G2992 was the largest and was associated with resistance to azithromycin, and with men who have sex with men (MSM), alongside G2400. G1407 and G2400 strains were related to high minimum inhibitory concentration (MICs) to cefixime and G1407 also to ceftriaxone. For the first time, G1861 and G2018 were described and associated with ciprofloxacin resistance and G2018 also with high MICs to ceftriaxone. CONCLUSION: Molecular typing is a useful tool to predict antimicrobial resistance. These results show the need to develop novel antimicrobials or to design new antimicrobial therapies based on drugs that show their efficacy against GI. This also highlights the importance of developing sexually transmitted infection (STI) surveillance in homosexual populations

INTRODUCCIÓN: el tratamiento de la gonorrea está amenazado por la resistencia antimicrobiana, y la disminución de la sensibilidad a las terapias recomendadas está emergiendo. Por ello la infección gonocócica (IG) se está convirtiendo en un problema de salud pública. MÉTODOS: la sensibilidad antimicrobiana se estudió por el método de difusión en disco, cefixima y ceftriaxona fueron testados por el método de dilución en agar y azitromicina por Etest. El genotipado se realizó por el método NG multi-antigen sequence typing (NG-MAST). Se definieron genogrupos con secuenciotipos (STs) relacionados. RESULTADOS: todas las cepas fueron sensibles a cefixima, ceftriaxona y gentamicina y el 1,8% resistentes a azitromicina. Se identificaron 531 STs y 6 genotipos (Gs) durante el período 2012-2015. El G2992 fue el más grande y se relacionó con resistencia a azitromicina, y con hombres que tienen sexo con hombres (HSH) junto con el G2400. Las cepas pertenecientes a los G1407 y G2400 se relacionaron con altas concentraciones mínimas inhibitorias (CMIs) a cefixima y el G1407 también a ceftriaxona. Se describe por primera vez la presencia del G1861 y G2018 y su relación con la resistencia a ciprofloxacino y la relación del G2018 con alta CMI a ceftriaxona. CONCLUSIÓN: El tipado molecular es una herramienta útil para predecir la resistencia antimicrobiana. Estos resultados muestran la necesidad de desarrollar nuevos antimicrobianos o nuevas terapias basadas en fármacos que demuestren su eficacia contra la IG. También muestra la importancia del desarrollo de la vigilancia de las infecciones de transmisión sexual (ITS) en la población homosexual

Epidemiological surveillance study of gonococcal infection in Northern Spain.

BACKGROUND: Treatment of gonorrhoea is threatened by antimicrobial resistance, and decreased susceptibility to recommended therapies is emerging. Thus, gonococcal infection (GI) is becoming a public health problem. The objectives of the present study were to monitor the antimicrobial sensitivity in Neisseria gonorrhoeae (NG) during 2011-2015 and to study their genogroups. METHODS: Antimicrobial susceptibility was studied by disc diffusion, in addition to the agar dilution method for cefixime and ceftriaxone and the Etest® for azithromycin. Genotyping was performed by the NG multi-antigen sequence typing (NG-MAST) method. Genogroups of closely related sequence types (STs) were defined. RESULTS: All the strains were susceptible to cefixime, ceftriaxone and gentamicin and 1.8% of the strains were resistant to azithromycin. A total of 531 STs and 6 genotypes (Gs) were identified during 2012-2015 period. G2992 was the largest and was associated with resistance to azithromycin, and with men who have sex with men (MSM), alongside G2400. G1407 and G2400 strains were related to high minimum inhibitory concentration (MICs) to cefixime and G1407 also to ceftriaxone. For the first time, G1861 and G2018 were described and associated with ciprofloxacin resistance and G2018 also with high MICs to ceftriaxone. CONCLUSION: Molecular typing is a useful tool to predict antimicrobial resistance. These results show the need to develop novel antimicrobials or to design new antimicrobial therapies based on drugs that show their efficacy against GI. This also highlights the importance of developing sexually transmitted infection (STI) surveillance in homosexual populations.