Scientific letter
A comparative evaluation of the sensitivity of one automated and one manual nucleic acid extraction methods for the performance of the Speed-oligo™ Direct Mycobacterium Tuberculosis assay
Evaluación comparativa de la sensibilidad de un método automatizado y uno manual para la extracción de ácidos nucleicos con el ensayo Speed-oligo™ Direct Mycobacterium Tuberculosis
María José Muñoz-Dávila
, María Aurora Cejudo García, Miguel José Martínez-Lirola
Corresponding author
Laboratorio de Microbiología, Unidad de Gestión Clínica de Biotecnología, Complejo Hospitalario Torrecárdenas, 04009 Almería, Spain
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] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Mercedes Treviño Castellano, Daniel Navarro de la Cruz, Paloma Areses Elizalde, Antonio Aguilera Guirao" "autores" => array:4 [ 0 => array:2 [ "nombre" => "Mercedes" "apellidos" => "Treviño Castellano" ] 1 => array:2 [ "nombre" => "Daniel" "apellidos" => "Navarro de la Cruz" ] 2 => array:2 [ "nombre" => "Paloma" "apellidos" => "Areses Elizalde" ] 3 => array:2 [ "nombre" => "Antonio" "apellidos" => "Aguilera Guirao" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0213005X14003206?idApp=UINPBA00004N" "url" => "/0213005X/0000003300000005/v2_201505140447/S0213005X14003206/v2_201505140447/en/main.assets" ] "en" => array:15 [ "idiomaDefecto" => true "cabecera" => "<span class="elsevierStyleTextfn">Scientific letter</span>" "titulo" => "A comparative evaluation of the sensitivity of one automated and one manual nucleic acid extraction methods for the performance of the Speed-oligo™ Direct Mycobacterium Tuberculosis assay" "tieneTextoCompleto" => true "saludo" => "Dear Editor," "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "360" "paginaFinal" => "361" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "María José Muñoz-Dávila, María Aurora Cejudo García, Miguel José Martínez-Lirola" "autores" => array:3 [ 0 => array:4 [ "nombre" => "María José" "apellidos" => "Muñoz-Dávila" "email" => array:2 [ 0 => "mjmd205@hotmail.com" 1 => "mariajose.munoz5@um.es" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">*</span>" "identificador" => "cor0005" ] ] ] 1 => array:2 [ "nombre" => "María Aurora" "apellidos" => "Cejudo García" ] 2 => array:2 [ "nombre" => "Miguel José" "apellidos" => "Martínez-Lirola" ] ] "afiliaciones" => array:1 [ 0 => array:2 [ "entidad" => "Laboratorio de Microbiología, Unidad de Gestión Clínica de Biotecnología, Complejo Hospitalario Torrecárdenas, 04009 Almería, Spain" "identificador" => "aff0005" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Evaluación comparativa de la sensibilidad de un método automatizado y uno manual para la extracción de ácidos nucleicos con el ensayo Speed-oligo™ Direct Mycobacterium Tuberculosis" ] ] "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">Nucleic acid amplification techniques are becoming widely used for mycobacterial detection as they combine high sensitivity, high specificity, and the rapid turnaround of results. Among the different steps that participate in ensuring a reliable molecular detection method, DNA extraction is crucial.</p><p id="par0010" class="elsevierStylePara elsevierViewall">The novel oligochromatographic assay, Speed-oligo™ Direct Mycobacterium Tuberculosis (SO-DMT) (Vircell S.L., Santa Fe, Granada, Spain), is based on PCR technology combined with a dipstick to detect the presence of <span class="elsevierStyleItalic">Mycobacterium</span> and specifically identify <span class="elsevierStyleItalic">Mycobacterium tuberculosis</span> complex (MTC) in clinical respiratory specimens.</p><p id="par0015" class="elsevierStylePara elsevierViewall">In our previous study, we demonstrated a high sensitivity of SO-DMT in detection of <span class="elsevierStyleItalic">Mycobacterium</span> spp. in acid fast bacilli (AFB) smear-positive clinical respiratory specimens and a simultaneous differentiation from MTC species.<a class="elsevierStyleCrossRef" href="#bib0005"><span class="elsevierStyleSup">1</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">In this study, we compared on performance of the SO-DMT assay, related to mycobacterial culture, by using two different nucleic acid extraction methods: the one manual included in the SO-DMT assay (Vircell-NXM) and another automated performed with the GXT DNA/RNA nucleic acid extraction kit (Hain Lifescience, Nehren, Germany) (Hain-ANXM).</p><p id="par0025" class="elsevierStylePara elsevierViewall">The purpose of comparison was, in case of a high correlation of results, to only perform the automated Hain-ANXM extraction for the SO-DMT assay with the aim of a single nucleic acid extraction with less hands-on-time which could also be used for the Genotype<span class="elsevierStyleSup">®</span> MTBDRplus test in case of MTC detection.</p><p id="par0030" class="elsevierStylePara elsevierViewall">SO-DMT and culture were used to prospectively assay 81 fresh respiratory specimens from 81 patients with suspicion of mycobacterial disease from August 2012 till June 2013. However, 23 could only be extracted by one method (21 by Vircell-NXM and 2 by Hain-ANXM). For this reason, they were excluded from the study as both extraction methods could not be compared. Also, 27 frozen respiratory specimens (culture results: MTC [12], and Nontuberculous mycobacteria (NTM) [15]) were retrospectively evaluated with the SO-DMT. Thus, a total of 85 specimens were included in the study. Respiratory specimens were decontaminated, examined by auramine staining according with a method previously published<a class="elsevierStyleCrossRef" href="#bib0010"><span class="elsevierStyleSup">2</span></a> and cultured following standard protocols.<a class="elsevierStyleCrossRefs" href="#bib0015"><span class="elsevierStyleSup">3,4</span></a> Generally, Mycobacteria were identified by GenoType™ Mycobacterium CM/AS assay (Hain Lifescience, Nehren, Germany) or BD MGIT™ TBc ID (Beckton Dickinson, USA). However, one NTM isolate included was finally identified as <span class="elsevierStyleItalic">Mycobacterium terrae</span> by sequence-based methods (Gregorio Marañon Hospital, Madrid (Spain)).</p><p id="par0035" class="elsevierStylePara elsevierViewall">Two aliquots of 650<span class="elsevierStyleHsp" style=""></span>μl of decontaminated specimens were maintained for a maximum of 7 days at 4<span class="elsevierStyleHsp" style=""></span>°C until assayed with SO-DMT. In those cases where we observed a disagreement among the results obtained by the two nucleic acid extraction methods, we confirmed the discrepancy by repeating the whole test again using new nucleic acid extracts from the beginning. In all cases, the disagreement kept. For the statistical analysis, the SPSS 15.0 software was used.</p><p id="par0040" class="elsevierStylePara elsevierViewall">Out of the 85 specimens, 81 (95.3%) were Acid-Fast Bacilli (AFB) smear-positive: 60 (70.6%) strong-positive (≥1–9 AFB/field [200×]) and 21 (24.7%) weak-positive (<1–9 AFB/field [200×]). Finally, 4 (4.7%) AFB negative and mycobacterial culture positive specimens (1 MTC and 3 NTM) were included. According to the culture results, 61 (71.8%) were positive for MTC and 24 (28.2% for NTM). The NTM species isolated were identified as <span class="elsevierStyleItalic">M. abscessus</span> (5 [5.9%]), <span class="elsevierStyleItalic">M. avium</span> (10 [11.8%]), <span class="elsevierStyleItalic">M. genavense</span> (1 [1.2%]), <span class="elsevierStyleItalic">M. gordonae</span> (1 [1.2%]), <span class="elsevierStyleItalic">M. intracellulare</span> (5 [5.9%]), <span class="elsevierStyleItalic">M. scrofulaceum</span> (1 [1.2%]), and <span class="elsevierStyleItalic">M. terrae</span> (1 [1.2%]).</p><p id="par0045" class="elsevierStylePara elsevierViewall">In the detection of <span class="elsevierStyleItalic">Mycobacterium</span> spp. in the 85 valuables specimens the Vircell-NXM showed an overall Sensitivity (S) of 0.87 (74/85) and Predictive Positive Value (PPV) of 1 (74/74); while Hain-ANXM showed a S of 0.8 (68/85) and PPV of 1 (68/68).</p><p id="par0050" class="elsevierStylePara elsevierViewall">When considering the subgroup of 81 specimens AFB smear-positive; the S of the SO-DMT Vircell nucleic acid extraction method was 0.9 (73/81), (S: 0.95 (57/60) in AFB strong-positives and S: 0.76 (16/21) in the AFB weak-positives); whereas with the Hain-ANXM, S was 0.83 (67/81) (S: 0.92 (55/60) in the AFB strong-positive and S: 0.57 (12/21) in the AFB weak-positive subgroup) (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>).</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0055" class="elsevierStylePara elsevierViewall">A total of 3 (2.9%) specimens gave a low detection signal for MTC (MTC +/−) while 2 (1.9%) specimens gave a low detection signal for NTM (NTM +/−) when Vircell-NXM was used. When Hain-ANXM was applied, a low detection signal for MTC (MTC +/−) and NTM (NTM +/−) was observed in 6 (5.8%) and 2 (1.9%) specimens respectively.</p><p id="par0060" class="elsevierStylePara elsevierViewall">None of the nucleic acid extraction methods used with the SO-DMT assay showed molecular detection of NTM in a MTC case or vice versa.</p><p id="par0065" class="elsevierStylePara elsevierViewall">In six specimens, four of them AFB weak-positive, no mycobacterial detection was observed with the automated extraction method. Meanwhile, in those six cases, the manual one showed a SO-DMT positive result (3 MTC and 3 NTM). But, in three out of these six cases (2 MTC and 1 NTM), the manual method gave a low detection signal. In accordance with previously exposed, our results showed a most efficient DNA extraction with the manual Vircell-NXM in comparison with the automated method. Specially, it happened in weakly positive AFB specimens. Moreover, the detection of mycobacterial DNA in AFB negative smear showed a very low S (S: 0.25) when we used any of those extraction methods. In any case, the SO-DMT assay showed a positive result only with the Hain-ANXM.</p><p id="par0070" class="elsevierStylePara elsevierViewall">The manual extraction method took approximately 1<span class="elsevierStyleHsp" style=""></span>h per specimen, which 15<span class="elsevierStyleHsp" style=""></span>min were hands-on time to be distributed across the whole time period. Whereas the automated extraction method took also 1<span class="elsevierStyleHsp" style=""></span>h per specimen, in which only 3<span class="elsevierStyleHsp" style=""></span>min were hands-on time at the beginning of the process.</p><p id="par0075" class="elsevierStylePara elsevierViewall">No previous study has compared the influences of two DNA extraction methods (a manual and an automated one) on the sensitivity of PCR amplification of the SO-DMT assay.</p><p id="par0080" class="elsevierStylePara elsevierViewall">The main advantage of the automated nucleic acid extraction method is that it only required 3<span class="elsevierStyleHsp" style=""></span>min hands-on-time, being the rest of the time fully automated.</p><p id="par0085" class="elsevierStylePara elsevierViewall">However, the S is more important than laboratory automatization or economy in the diagnosis of tuberculosis. The automated system missed two specimens even among strong positive specimens.</p><p id="par0090" class="elsevierStylePara elsevierViewall">There is a limitation in our study as we have not compared the manual and the automated extraction method with the SO-DMT in another kind of specimen different from the respiratory one.</p><p id="par0095" class="elsevierStylePara elsevierViewall">Further studies will be necessary to assess the efficiency of both extraction methods with the SO-DMT assay when using specimens not belonging to the respiratory tract in which mycobacterial isolates have also been described.</p><p id="par0100" class="elsevierStylePara elsevierViewall">In conclusion, even though the automated extraction method requires only 3<span class="elsevierStyleHsp" style=""></span>min hands-on-time, it should never be used alone, without the manual one, neither in strong positive nor in weak positive auramine staining specimens.</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0005">Conflict of interest</span><p id="par0110" class="elsevierStylePara elsevierViewall">None of the authors have conflict of interest.</p></span></span>" "textoCompletoSecciones" => array:1 [ "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0005" "titulo" => "Conflict of interest" ] 1 => array:2 [ "identificador" => "xack164538" "titulo" => "Acknowledgement" ] 2 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "multimedia" => array:1 [ 0 => array:7 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:2 [ "leyenda" => "<p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">PPV: positive predictive value.</p>" "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="table-head " align="left" valign="top" scope="col">Extraction method \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " colspan="5" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Specimens</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">All(<span class="elsevierStyleItalic">N</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>85)<span class="elsevierStyleItalic">n</span>/<span class="elsevierStyleItalic">N</span> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">AFB+(<span class="elsevierStyleItalic">N</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>81)<span class="elsevierStyleItalic">n</span>/<span class="elsevierStyleItalic">N</span> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">AFB−(<span class="elsevierStyleItalic">N</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>4)<span class="elsevierStyleItalic">n</span>/<span class="elsevierStyleItalic">N</span> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">MTC(<span class="elsevierStyleItalic">N</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>61)<span class="elsevierStyleItalic">n</span>/<span class="elsevierStyleItalic">N</span> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">NTM(<span class="elsevierStyleItalic">N</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>24)<span class="elsevierStyleItalic">n</span>/<span class="elsevierStyleItalic">N</span> \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry " colspan="6" align="left" valign="top"><span class="elsevierStyleItalic">Vircell-NXM</span></td></tr><tr title="table-row"><td class="td" title="table-entry " rowspan="2" align="left" valign="top"><span class="elsevierStyleHsp" style=""></span>Sensitivity</td><td class="td" title="table-entry " align="char" valign="top">0.87 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.90 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.96 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.62 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">74/85 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">73/81 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1/4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">59/61 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">15/24 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " rowspan="2" align="left" valign="top"><span class="elsevierStyleHsp" style=""></span>PPV</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">74/74 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">57/57 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1/1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">59/59 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">15/15 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " colspan="6" align="left" valign="top"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="table-entry " colspan="6" align="left" valign="top"><span class="elsevierStyleItalic">Hain-ANXM</span></td></tr><tr title="table-row"><td class="td" title="table-entry " rowspan="2" align="left" valign="top"><span class="elsevierStyleHsp" style=""></span>Sensitivity</td><td class="td" title="table-entry " align="char" valign="top">0.8 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.83 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.25 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.92 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">68/85 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">67/81 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1/4 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">56/61 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">12/24 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " rowspan="2" align="left" valign="top"><span class="elsevierStyleHsp" style=""></span>PPV</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="char" valign="top">68/68 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">67/67 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">1/1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">56/56 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">12/12 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab804774.png" ] ] ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Sensitivity and positive predictive values of SO-DMT assay compared to culture with both nucleic acid extraction methods studied.</p>" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:4 [ 0 => array:3 [ "identificador" => "bib0005" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Evaluation of the Speed-oligo Direct Mycobacterium tuberculosis assay for molecular detection of mycobacteria in clinical respiratory specimens" "autores" => array:1 [ 0 => array:2 [ "etal" => true "autores" => array:6 [ 0 => "A. Lara-Oya" 1 => "P. Mendoza-Lopez" 2 => "J. Rodriguez-Granger" 3 => "A.M. Fernández-Sánchez" 4 => "M.P. Bermúdez-Ruiz" 5 => "I. Toro-Peinado" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1128/JCM.00533-12" "Revista" => array:6 [ "tituloSerie" => "J Clin Microbiol" "fecha" => "2013" "volumen" => "51" "paginaInicial" => "77" "paginaFinal" => "82" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23100355" "web" => "Medline" ] ] ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bib0010" "etiqueta" => "2" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "An ultraviolet-fixed thick-blotch preparation improves sensitivity of auramine staining" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "M.A. Cejudo" 1 => "M.J. Munoz-Davila" 2 => "M.J. Martinez-Lirola" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1128/JCM.00974-13" "Revista" => array:6 [ "tituloSerie" => "J Clin Microbiol" "fecha" => "2013" "volumen" => "51" "paginaInicial" => "3400" "paginaFinal" => "3402" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/23863571" "web" => "Medline" ] ] ] ] ] ] ] ] 2 => array:3 [ "identificador" => "bib0015" "etiqueta" => "3" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Sputum digestion and decontamination with N-acetyl-<span class="elsevierStyleSmallCaps">l</span>-cysteine-sodium hydroxide for culture of mycobacteria" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "G.P. Kubica" 1 => "W.E. Dye" 2 => "M.L. Cohn" 3 => "G. Middlebrook" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1164/arrd.1963.87.5.775" "Revista" => array:6 [ "tituloSerie" => "Am Rev Respir Dis" "fecha" => "1963" "volumen" => "87" "paginaInicial" => "775" "paginaFinal" => "779" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/13927224" "web" => "Medline" ] ] ] ] ] ] ] ] 3 => array:3 [ "identificador" => "bib0020" "etiqueta" => "4" "referencia" => array:1 [ 0 => array:1 [ "referenciaCompleta" => "Pfyffer G, Frantiska P. <span class="elsevierStyleItalic">Mycobacterium</span>: general characteristics, laboratory detection, and staining procedures. In: Carroll KC, Versalovic J, Funke G, Jorgensen JH, Landry ML, Warnock D, editors. Manual of clinical microbiology. 10th ed. Washington, DC:American Society of Microbiology; 2011." ] ] ] ] ] ] ] "agradecimientos" => array:1 [ 0 => array:4 [ "identificador" => "xack164538" "titulo" => "Acknowledgement" "texto" => "<p id="par0105" class="elsevierStylePara elsevierViewall">We are grateful to Pablo Mendoza (Vircell S.L., Santa Fe, Granada, Spain) for providing the Speed-oligo™ Direct Mycobacterium Tuberculosis kits for the study.</p>" "vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/0213005X/0000003300000005/v2_201505140447/S0213005X14002985/v2_201505140447/en/main.assets" "Apartado" => array:4 [ "identificador" => "15253" "tipo" => "SECCION" "es" => array:2 [ "titulo" => "Cartas científicas" "idiomaDefecto" => true ] "idiomaDefecto" => "es" ] "PDF" => "https://static.elsevier.es/multimedia/0213005X/0000003300000005/v2_201505140447/S0213005X14002985/v2_201505140447/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0213005X14002985?idApp=UINPBA00004N" ]
| Year/Month | Html | Total | |
|---|---|---|---|
| 2024 October | 6 | 7 | 13 |
| 2024 September | 16 | 9 | 25 |
| 2024 August | 14 | 14 | 28 |
| 2024 July | 8 | 10 | 18 |
| 2024 June | 13 | 7 | 20 |
| 2024 May | 8 | 13 | 21 |
| 2024 April | 14 | 2 | 16 |
| 2024 March | 18 | 3 | 21 |
| 2024 February | 16 | 4 | 20 |
| 2024 January | 27 | 7 | 34 |
| 2023 December | 15 | 7 | 22 |
| 2023 November | 24 | 17 | 41 |
| 2023 October | 35 | 18 | 53 |
| 2023 September | 20 | 1 | 21 |
| 2023 August | 18 | 6 | 24 |
| 2023 July | 19 | 11 | 30 |
| 2023 June | 13 | 5 | 18 |
| 2023 May | 16 | 1 | 17 |
| 2023 April | 11 | 5 | 16 |
| 2023 March | 20 | 7 | 27 |
| 2023 February | 7 | 6 | 13 |
| 2023 January | 11 | 11 | 22 |
| 2022 December | 16 | 14 | 30 |
| 2022 November | 29 | 17 | 46 |
| 2022 October | 19 | 15 | 34 |
| 2022 September | 22 | 20 | 42 |
| 2022 August | 36 | 7 | 43 |
| 2022 July | 21 | 11 | 32 |
| 2022 June | 20 | 6 | 26 |
| 2022 May | 25 | 9 | 34 |
| 2022 April | 43 | 9 | 52 |
| 2022 March | 30 | 11 | 41 |
| 2022 February | 13 | 4 | 17 |
| 2022 January | 16 | 13 | 29 |
| 2021 December | 14 | 20 | 34 |
| 2021 November | 15 | 11 | 26 |
| 2021 October | 17 | 13 | 30 |
| 2021 September | 13 | 17 | 30 |
| 2021 August | 12 | 9 | 21 |
| 2021 July | 25 | 14 | 39 |
| 2021 June | 20 | 10 | 30 |
| 2021 May | 16 | 12 | 28 |
| 2021 April | 42 | 14 | 56 |
| 2021 March | 19 | 22 | 41 |
| 2021 February | 17 | 8 | 25 |
| 2021 January | 26 | 25 | 51 |
| 2020 December | 17 | 14 | 31 |
| 2020 November | 15 | 22 | 37 |
| 2020 October | 14 | 11 | 25 |
| 2020 September | 34 | 8 | 42 |
| 2020 August | 20 | 9 | 29 |
| 2020 July | 17 | 8 | 25 |
| 2020 June | 8 | 16 | 24 |
| 2020 May | 26 | 11 | 37 |
| 2020 April | 5 | 8 | 13 |
| 2020 March | 14 | 6 | 20 |
| 2020 February | 15 | 13 | 28 |
| 2020 January | 18 | 11 | 29 |
| 2019 December | 26 | 16 | 42 |
| 2019 November | 9 | 12 | 21 |
| 2019 October | 13 | 8 | 21 |
| 2019 September | 8 | 5 | 13 |
| 2019 August | 12 | 6 | 18 |
| 2019 July | 18 | 13 | 31 |
| 2019 June | 28 | 16 | 44 |
| 2019 May | 73 | 49 | 122 |
| 2019 April | 35 | 16 | 51 |
| 2019 March | 7 | 8 | 15 |
| 2019 February | 14 | 12 | 26 |
| 2019 January | 8 | 5 | 13 |
| 2018 December | 7 | 4 | 11 |
| 2018 November | 11 | 6 | 17 |
| 2018 October | 21 | 5 | 26 |
| 2018 September | 6 | 4 | 10 |
| 2018 August | 10 | 3 | 13 |
| 2018 July | 8 | 0 | 8 |
| 2018 June | 6 | 1 | 7 |
| 2018 May | 8 | 2 | 10 |
| 2018 April | 8 | 3 | 11 |
| 2018 March | 3 | 2 | 5 |
| 2018 February | 4 | 2 | 6 |
| 2018 January | 11 | 3 | 14 |
| 2017 December | 1 | 2 | 3 |
| 2017 November | 16 | 8 | 24 |
| 2017 October | 6 | 3 | 9 |
| 2017 September | 11 | 8 | 19 |
| 2017 August | 14 | 4 | 18 |
| 2017 July | 6 | 6 | 12 |
| 2017 June | 15 | 6 | 21 |
| 2017 May | 17 | 4 | 21 |
| 2017 April | 12 | 3 | 15 |
| 2017 March | 15 | 55 | 70 |
| 2017 February | 9 | 2 | 11 |
| 2017 January | 12 | 0 | 12 |
| 2016 December | 19 | 3 | 22 |
| 2016 November | 27 | 3 | 30 |
| 2016 October | 53 | 5 | 58 |
| 2016 September | 34 | 5 | 39 |
| 2016 August | 21 | 1 | 22 |
| 2016 July | 25 | 2 | 27 |
| 2016 June | 24 | 6 | 30 |
| 2016 May | 14 | 1 | 15 |
| 2016 April | 31 | 33 | 64 |
| 2016 March | 21 | 12 | 33 |
| 2016 February | 19 | 15 | 34 |
| 2016 January | 24 | 13 | 37 |
| 2015 December | 31 | 16 | 47 |
| 2015 November | 34 | 12 | 46 |
| 2015 October | 32 | 18 | 50 |
| 2015 September | 29 | 10 | 39 |
| 2015 August | 40 | 6 | 46 |
| 2015 July | 35 | 13 | 48 |
| 2015 June | 41 | 15 | 56 |
| 2015 May | 91 | 53 | 144 |
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