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dc.contributor.authorSanoussi, C.N.
dc.contributor.authorde Jong, B.C.
dc.contributor.authorAffolabi, D.
dc.contributor.authorMeehan, Conor J.
dc.contributor.authorOdoun, M.
dc.contributor.authorRigouts, L.
dc.date.accessioned2019-09-16T21:29:21Z
dc.date.accessioned2019-09-30T15:29:55Z
dc.date.available2019-09-16T21:29:21Z
dc.date.available2019-09-30T15:29:55Z
dc.date.issued2019-07
dc.identifier.citationSanoussi CN, de Jong BC, Affolabi D et al (2019) Storage of Sputum in Cetylpyridinium Chloride, OMNIgene.SPUTUM, and Ethanol Is Compatible with Molecular Tuberculosis Diagnostic Testing. Journal of Clinical Microbiology. 57(7): e00275-19.en_US
dc.identifier.urihttp://hdl.handle.net/10454/17275
dc.descriptionYesen_US
dc.description.abstractWe compared cetylpyridinium chloride (CPC), ethanol (ETOH), and OMNIgene.SPUTUM (OMNI) for 28-day storage of sputum at ambient temperature before molecular tuberculosis diagnostics. Three sputum samples were collected from each of 133 smear-positive tuberculosis (TB) patients (399 sputum samples). Each patient's sputum was stored with either CPC, ETOH, or OMNI for 28 days at ambient temperature, with subsequent rpoB amplification targeting a short fragment (81 bp, GeneXpert MTB/RIF [Xpert]) or a long fragment (1,764 bp, in-house nested PCR). For 36 patients, Xpert was also performed at baseline on all 108 fresh sputum samples. After the 28-day storage (D28), Xpert positivity did not significantly differ between storage methods. In contrast, higher positivity for rpoB nested PCR was obtained with OMNI (n = 125, 94%) than with ETOH (n = 114, 85.7%; P = 0.001). Smears with scanty acid-fast bacilli (AFB) had lower rpoB PCR positivity with ETOH storage (n = 10, 41.7%) than with CPC (n = 16, 66.7%; difference, 25%; 95% confidence interval [CI], 3.5 to 46.5; P = 0.031) or OMNI (n = 16, 69.6%; difference, 26.1%; 95% CI, 3.8 to 48.4; P = 0.031), with no difference between CPC and OMNI. Poststorage, the threshold cycle (CT ) values significantly decreased compared to those prestorage with ETOH (difference, -1.1; 95% CI, -1.6 to -0.6; P = 0.0001) but not with CPC (P = 0.915) or OMNI (P = 0.33). For one patient's ETOH- and CPC-stored specimens with a CT of <10, Xpert gave results of rifampin false resistant at D28, which was resolved by repeating Xpert on a 1/100 diluted specimen. In conclusion, 28-day storage of sputum in OMNI, CPC, or ETOH at ambient temperature does not impact short-fragment PCR (Xpert), including for low smear grades. However, for long-fragment PCR, ETOH yielded a lower PCR positivity for low smear grades, while the performance of OMNI and CPC was excellent for all smear grades. (The study has been registered at ClinicalTrials.gov under registration number NCT02744469.).en_US
dc.description.sponsorshipDirectorate General for Development (DGD), Belgium (FA4 to C.N.S., B.C.D.J., D.A., and L.R.), and the European Research Council-INTERRUPTB starting grant (number 311725 to B.C.D.J., C.J.M., and L.R.)en_US
dc.language.isoenen_US
dc.relation.isreferencedbyhttps://doi.org/10.1128/JCM.00275-19en_US
dc.rights(c) 2019 The Authors. This is an Open Access article distributed under the Creative Commons CC-BY license (https://creativecommons.org/licenses/by/4.0/)en_US
dc.subjectAFB scantyen_US
dc.subjectOMNIgene.SPUTUMen_US
dc.subjectXperten_US
dc.subjectCetylpyridinium chlorideen_US
dc.subjectEthanolen_US
dc.subjectIsolateen_US
dc.subjectMolecular testsen_US
dc.subjectShort/long-fragment PCRen_US
dc.subjectSputumen_US
dc.subjectStorageen_US
dc.titleStorage of Sputum in Cetylpyridinium Chloride, OMNIgene.SPUTUM, and Ethanol Is Compatible with Molecular Tuberculosis Diagnostic Testingen_US
dc.status.refereedYesen_US
dc.date.Accepted2019-05-05
dc.date.application2019-05-15
dc.typeArticleen_US
dc.type.versionPublished versionen_US
dc.date.updated2019-09-16T20:29:22Z
refterms.dateFOA2019-09-30T15:30:19Z


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