Improved Performance of a Rapid Immunochromatographic Assay for Detection of PBP2a in Non-Staphylococcus aureus Staphylococcal Species

Canver, Matthew C.; Gonzalez, Mark D.; Ford, Bradley; Arnold, Amanda R.; Lawhon, Sara D.; Burnham, Carey‐Ann D.; Jenkins, Stephen G.; Burd, Eileen M.; Westblade, Lars F.

doi:10.1128/jcm.01417-18

Cited by 14 publications

(11 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The PBP2a SA culture colony test, which is approved by the FDA for PBP2a detection in S. aureus, was 100% sensitive and 100% specific for S. epidermidis isolates by the use of the default manufacturer instructions (i.e., without oxacillin induction). These findings are consistent with other studies that reported 100% sensitivity and specificity for other staphylococcal species, including 50 S. epidermidis (11) and 54 S. schleiferi (8) isolates. Although the PBP2= latex agglutination test kit is FDA approved for both S. aureus and CoNS (with oxacillin induction), two false negatives were identified, resulting in overall sensitivity and specificity of 95.8% and 100%, respectively.…”

Section: Discussionsupporting

confidence: 93%

Evaluation of Oxacillin and Cefoxitin Disk Diffusion and Microbroth Dilution Methods for Detecting mecA -Mediated β-Lactam Resistance in Contemporary Staphylococcus epidermidis Isolates

et al. 2019

Self Cite

View full text Add to dashboard Cite

Methicillin (β-lactam) resistance in Staphylococcus epidermidis is mediated by the mecA gene, with resistance reported to be as high as 90%. The goal of this study was to evaluate oxacillin and cefoxitin disk diffusion (DD) and broth microdilution (BMD) methods for the detection of mecA-mediated β-lactam resistance in 100 human isolates of S. epidermidis (48 mecA-positive isolates and 52 mecA negative isolates). Oxacillin DD tests using the Clinical and Laboratory Standards Institute (CLSI) M100-S28 breakpoints for S. pseudintermedius/S. schleiferi accurately differentiated mecA-positive and -negative S. epidermidis isolates, with categorical agreement (CA) of 100% and no very major errors (VMEs) or major errors (MEs) identified. Likewise, oxacillin BMD and cefoxitin DD tests using the coagulase-negative Staphylococcus species (CoNS) breakpoints were highly reliable for detecting mecA-mediated β-lactam resistance in S. epidermidis isolates. For cefoxitin DD and BMD results interpreted using S. aureus/S. lugdunensis breakpoints, the CA was 97.6% and 96.2%, respectively. There were 4.9% VMEs for cefoxitin DD with 0% MEs, and 3.6% VMEs and 3.9% MEs for cefoxitin BMD. Oxacillin BMD using S. aureus/S. lugdunensis breakpoints yielded the highest VMEs at 17.4% and 90% CA. Our findings demonstrate that oxacillin DD tests using the CLSI M100-S28 breakpoints for S. pseudintermedius/S. schleiferi and oxacillin BMD and cefoxitin DD tests using the CoNS breakpoints reliably identified mecA-mediated β-lactam resistance in S. epidermidis. Using mecA PCR as the gold standard, the PBP2a SA culture colony test (Abbott Diagnostics) exhibited 100% sensitivity and specificity whereas 2 false negatives were identified using the PBP2ʹ latex agglutination test kit (Thermo Fisher Scientific) with sensitivity and specificity of 95.8% and 100%, respectively.

show abstract

Section: Discussionsupporting

confidence: 93%

Evaluation of Oxacillin and Cefoxitin Disk Diffusion and Microbroth Dilution Methods for Detecting mecA -Mediated β-Lactam Resistance in Contemporary Staphylococcus epidermidis Isolates

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…Of note, the red lines observed with methicillin-resistant SNA isolates were often less intense than those observed with MRSA isolates, indicating that the operator must strictly respect the inoculum recommendations of the manufacturer (one heaped 1-l bacteriological loop of colonies) to avoid false-negative results. The high sensitivity for mecA-positive isolates is concordant with the results obtained with the previous version for S. aureus isolates (2)(3)(4)9), and our study confirmed, using a large collection of species and isolates, that the performance of this revised assay was largely improved for SNA species, as reported by Canver et al (6). Conversely, unlike with the previous version of the assay, no improvement in the detection of mecCpositive MRSA isolates was observed with cefoxitin induction, indicating that the antibodies and/or protocol used in the revised assay no longer allow the detection of PBP2c.…”

supporting

confidence: 92%

Performance of the Revised Version of an Immunochromatographic Assay for Detection of mecA - and mecC -Mediated Methicillin Resistance in Staphylococci

et al. 2019

View full text Add to dashboard Cite

D etection of methicillin resistance in staphylococcal isolates is crucial to quickly optimize treatment in cases of severe staphylococcal infections. Rapid genetic or phenotypic tests have been developed to detect isolates harboring the mecA gene, which is responsible for methicillin resistance related to additional penicillin-binding protein 2a (PBP2a) expression. The discovery in 2011 of a mecA variant, named mecC, encoding PBP2c and showing only 70% nucleotide sequence homology, was a real challenge for laboratories, since most of the tests targeting mecA failed to detect mecC (1). The first version of the immunochromatographic assay developed by Alere (PBP2a culture colony test; Alere, Scarborough, ME), based on blue-colored monoclonal antibodies targeting PBP2a, has been widely evaluated. It demonstrated 100% specificity and sensitivity for mecA-positive Staphylococcus aureus strains but displayed lower sensitivity for Staphylococcus non-aureus (SNA) species such as Staphylococcus epidermidis (2-4). For mecC-positive isolates, this assay revealed a lack of sensitivity unless PBP2c expression was induced by 18-h culture around a cefoxitin disk, which increased the sensitivity from 8.7% to 100% (5). Alere recently updated this test, whose intended use is still the detection of PBP2a from S. aureus isolates, in order to allow storage at room temperature and changed the antibodies immobilized on the nitrocellulose membrane, which are now recombinant monoclonal antibody fragments with a conjugate labeled red (6). Here, we evaluated the revised version of the assay, the PBP2a SA culture colony test (SACCT) (Alere), using a collection of both S. aureus and SNA isolates, in order to determine whether the technical changes, particularly the use of antibody fragments, affected the performance of the immunochromatographic test. A large collection of methicillin-susceptible S. aureus and methicillin-resistant S. aureus (MRSA) isolates (n ϭ 83) belonging to more than 50 different clones and methicillin-susceptible and methicillin-resistant SNA isolates (n ϭ 122) belonging to 14 different species was tested (see Table 1 for details). The presence of mecA and mecC was confirmed using PCR (7, 8). All S. aureus isolates had been previously characterized by spa typing, multilocus sequence typing, and/or microarray analysis (Alere Technologies, Jena, Germany) to ensure diversity of the tested strains, which were representative of the most prevalent clones circulating in Europe. Isolates were grown for 18 to 24 h on Columbia sheep blood agar plates (bioMérieux, La-Balme-les-Grottes, France) and then were tested directly using the PBP2a SACCT, according to the manufacturer's instructions. The assay was also performed after PBP2a/PBP2c induction using colonies harvested around a cefoxitin disk (30 g). The testing was performed by a single

show abstract

“…ELISAs eliminate sophisticated sample pretreatment and can tolerate certain matrix interferences because of washes between steps. Compared with ELISAs, immunochromatographic lateral flow strip test (ILFST) is simpler and rapid immunoassay that has been used in detecting pesticides (Byzova, Urusov, Zherdev, & Dzantiev, 2018;Ismail et al, 2016), veterinary drugs (Chen et al, 2017;Shelver & Smith, 2018), mycotoxins (Di Nardo et al, 2019;Majdinasab, Zareian, Zhang, & Li, 2019), bacteria (Canver et al, 2019;Saleh, Göttig, & Hamprecht, 2018), and viruses (Lyoo et al, 2017;Senthilkumaran et al, 2017). ILFST integrates all the required reagents into itself and can provide the results within 10 min that can be determined both qualitatively and quantitatively with unaided eyes and a strip reader, respectively.…”

Section: Introductionmentioning

confidence: 99%

Development of an immunochromatographic lateral flow strip test for the rapid detection of diclofenac in medicinal wine

Yang

Wang

Yang

et al. 2020

Food and Agricultural Immunology

View full text Add to dashboard Cite

A rapid immunochromatographic lateral flow strip test (ILFST) of competitive format for the detection of diclofenac (DCF) in medicinal wine samples was developed. The half-inhibitory concentration (IC 50) values of the ILFST for DCF and the limit of detection (LOD) in DCF-spiked samples were 0.66 and 0.15 ng/mL, respectively. Recoveries of DCF from spiked medicinal wine samples were from 84.4% to 85.9% within an assay (intra-assay) and from 83.7% to 88.4% between assays (inter-assay). The coefficients of variation (CV) for intra-assay and inter-assay were 4.10-7.56% and 4.71-8.94%, respectively. These results indicated that the ILFST was highly sensitive and reproducible. The same authentic medicinal wine samples were tested by both ILFST and high-performance liquid chromatography (HPLC) respectively. The results showed no significant differences between the two methods. Therefore, the ILFST can be widely used as a convenient tool for both qualitative and quantitative detection of DCF residues in medicinal wine samples.

show abstract

Improved Performance of a Rapid Immunochromatographic Assay for Detection of PBP2a in Non-Staphylococcus aureus Staphylococcal Species

Cited by 14 publications

References 14 publications

Evaluation of Oxacillin and Cefoxitin Disk Diffusion and Microbroth Dilution Methods for Detecting mecA -Mediated β-Lactam Resistance in Contemporary Staphylococcus epidermidis Isolates

Evaluation of Oxacillin and Cefoxitin Disk Diffusion and Microbroth Dilution Methods for Detecting mecA -Mediated β-Lactam Resistance in Contemporary Staphylococcus epidermidis Isolates

Performance of the Revised Version of an Immunochromatographic Assay for Detection of mecA - and mecC -Mediated Methicillin Resistance in Staphylococci

Development of an immunochromatographic lateral flow strip test for the rapid detection of diclofenac in medicinal wine

Contact Info

Product

Resources

About