Sensitivity and specificity of indirect ELISA for the detection of antibody titers against BVDV from beef cattle raised in Pará State

The aims of this study were to establish the prevalence of anti-bovine viral diarrhea virus (BVDV) antibodies (Ab) in beef cattle raised in Pará state, to compare the prevalence of seropositive animals to BVDV using a commercial indirect enzyme-linked immunosorbent assay kit (iELISA) and the virus neutralization (VN) test, and finally, to determine the sensitivity (Se) and specificity (Sp) of the iELISA for the detection of anti-BVDV Ab using VN as a gold standard. A total of 400 serum blood samples from Nelore cows aged at least 24 months from five farms in the Pará state from two mesoregions (Metropolitan Region of Belem and Northeast of Pará) were analyzed. All animals were vaccinated against brucellosis and foot-and-mouth disease. The examination of anti-BVDV Ab with VN was performed in the Laboratory of Bovine Viruses of the Biological Institute of Sao Paulo as described in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. For VN, bovine kidney epithelial cells from the Madin Darby Bovine Kidney (MDBK) strain were used. The determinations of anti-BVDV Ab were performed with the iELISA test at the Laboratory of Immunology and Microbiology of the Federal Rural University of Amazonia according to the manufacturer's recommendations. The results were classified as follows: (a) correct positive diagnosis, (b) incorrect positive diagnosis, (c) correct negative diagnosis, and (d) incorrect negative diagnosis, according to the results obtained from VN. From the values obtained from VN and iELISA, Se [(a ÷ a + d) × 100], Sp [(c ÷ c + b) × 100], positive predictive value [(a ÷ a + B) × 100], and negative predictive value [(c ÷ c + d) × 100] were calculated for iELISA. The frequencies (%) of seropositive animals were determined and compared both between the different tests (iELISA and VN) and between the different farms (1, 2, 3, 4, and 5). The statistical analysis was performed with a significance level of 5%. The prevalence of seropositive animals was found to be different (P < 0.0001) using VN (39.25% [157/400]) and iELISA (54.50% [218/400]). It was observed that the Se and Sp of the iELISA assay were 98.72% and 74.07%, respectively. Of the total, 25.93% (63/243) of the samples were considered false-positive and 1.27% false-negative (2/157). It was concluded that the BVDV infection is present in beef cattle herds of the state of Para. Based on the speed of execution, ease of handling, and high Se of the iELISA, it is suggested that this assay can be used as a screening test for the detection of anti-BVDV Ab with the aim of eliminating infected animals from large herds of beef cattle.

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Indirect ELISA (iELISA) standardization for the diagnosis of bovine enzootic leukosis

Pesquisa Veterinária Brasileira ◽

10.1590/1678-5150-pvb-6731 ◽

2020 ◽

Vol 40 (12) ◽

pp. 977-984

Author(s):

Cláudia F. Resende ◽

Grazielle C. F. Galinari ◽

Raphael M. Victor ◽

Telissa C. Kassar ◽

Thiago L.M. Arcebispo ◽

...

Keyword(s):

Predictive Value ◽

Indirect Elisa ◽

Leukemia Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Kappa Index ◽

Tadarida Brasiliensis ◽

Diarrhea Virus ◽

Bovine Leukosis ◽

Specific Reactions ◽

Serum Elisa

ABSTRACT: Enzootic bovine leukosis (EBL) is an infectious disease caused by bovine leukemia virus (BLV) that affects cattle worldwide. Agar gel immunodiffusion (AGID) was the reference test for EBL diagnosis for many years, but enzyme-linked immunosorbent assay (ELISA) showed higher sensitivity, was faster to perform, and resulted in an objective reading. However, the importation of ELISA kits is lengthy and expensive, and currently, no AGID kits are available in Brazil. The aim of this work was to standardize an indirect ELISA (iELISA) for EBL diagnosis using BLV antigens produced in Tadarida brasiliensis lung (Tb1Lu) cells, which are Bovine viral diarrhea virus (BVDV) free, unlike fetal lamb kidney (FLK) cells, currently used for this purpose. Following standardization, iELISA results were compared with those obtained by AGID and the commercial Chekit Leucose-Serum ELISA. Compared to AGID, iELISA had 94,44% sensitivity, 75.68% specificity, 79.10% positive predictive value (PPV) and 93.30% negative predictive value (NPV), with 84% concordance and a Kappa index of 0.699. Compared to the Chekit Leucose-Serum ELISA, iELISA showed 92.60% sensitivity, 87.09% specificity, 90.27% PPV and 90,00% NPV, with 90.27% concordance and a Kappa index of 0.801. Taking into account the high agreement with the traditional tests and the absence of non-specific reactions with BVDV, the developed assay could be used as diagnostic method to control EBL in Brazil.

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Accuracy of the dental pulp sensibility test using cold spray for the diagnosis of pulp diseases: an observational clinical study

Bioscience Journal ◽

10.14393/bj-v37n0a2021-60365 ◽

2021 ◽

Vol 37 ◽

pp. e37040

Author(s):

Felipe de Souza Matos ◽

Thaís Christina Cunha ◽

Maria Amália Gonzaga Ribeiro ◽

Camila Silva Araujo ◽

Ítalo de Macedo Bernardino ◽

...

Keyword(s):

Cold Spray ◽

Predictive Value ◽

Dental Pulp ◽

False Negative ◽

Cold Test ◽

Chi Square ◽

True Negative ◽

Cross Sectional ◽

Significance Level ◽

Clinical Variables

The dental pulp sensibility test is one of the main auxiliary resources for the diagnosis of pulp pathologies, and its accuracy is still debatable. This cross-sectional observational study evaluated the accuracy of the pulp sensibility test (PST) using cold spray (1,1,1,2-tetrafluoroethane) for the diagnosis of pulp diseases and determined the effect of individual and clinical variables on the reliability of this test. The paper was designed following the STROBE statement. Sixty patients with indications for primary endodontic treatment were selected and examined from August 2017 to July 2018. Data collection was performed through interviews, clinical/radiographic examinations and the PST. The results of the cold test, along with data on sex, age, the tooth type regarding the root number, and the presence of restorations and caries, as well as the recent consumption of analgesics, were recorded. The presence of bleeding within the pulp chamber was used as the gold standard to compare with the clinical diagnosis and to identify the true-positive, false-positive, true-negative, and false-negative responses. The accuracy of PST achieved in subgroups of individual and clinical variables was compared using the chi-square test with a significance level of 5% (p < 0.05). The PST with the use of cold spray showed a sensitivity of 0.88, a speciﬁcity of 1.00, a positive predictive value of 1.00, a negative predictive value of 0.86, and an accuracy of 0.93. The accuracy of the cold spray was not affected by individual or clinical variables. The PST with the use of cold spray is an accurate and reliable method for determining the diagnosis of pulp diseases, especially in cases of pulp vitality or irreversible pulpitis.

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Tissue plasminogen activator as a novel diagnostic aid in acute pulmonary embolism

VASA ◽

10.1024/0301-1526/a000392 ◽

2014 ◽

Vol 43 (6) ◽

pp. 450-458 ◽

Cited By ~ 1

Author(s):

Julio Flores ◽

Ángel García-Avello ◽

Esther Alonso ◽

Antonio Ruíz ◽

Olga Navarrete ◽

...

Keyword(s):

Pulmonary Embolism ◽

Negative Predictive Value ◽

Plasminogen Activator ◽

Tissue Plasminogen Activator ◽

Predictive Value ◽

Acute Pulmonary Embolism ◽

Enzyme Linked Immunosorbent Assay ◽

Diagnostic Utility ◽

Tissue Plasminogen ◽

D Dimer

Background: We evaluated the diagnostic efficacy of tissue plasminogen activator (tPA), using an enzyme-linked immunosorbent assay (ELISA) and compared it with an ELISA D-dimer (VIDAS D-dimer) in acute pulmonary embolism (PE). Patients and methods: We studied 127 consecutive outpatients with clinically suspected PE. The diagnosis of PE was based on a clinical probability pretest for PE and a strict protocol of imaging studies. A plasma sample to measure the levels of tPA and D-dimer was obtained at enrollment. Diagnostic accuracy for tPA and D-dimer was determined by the area under the receiver operating characteristic (ROC) curve. Sensitivity, specificity, predictive values, and the diagnostic utility of tPA with a cutoff of 8.5 ng/mL and D-dimer with a cutoff of 500 ng/mL, were calculated for PE diagnosis. Results: PE was confirmed in 41 patients (32 %). Areas under ROC curves were 0.86 for D-dimer and 0.71 for tPA. The sensitivity/negative predictive value for D-dimer using a cutoff of 500 ng/mL, and tPA using a cutoff of 8.5 ng/mL, were 95 % (95 % CI, 88–100 %)/95 % (95 % CI, 88–100 %) and 95 % (95 % CI, 88–100 %)/94 %), respectively. The diagnostic utility to exclude PE was 28.3 % (95 % CI, 21–37 %) for D-dimer and 24.4 % (95 % CI, 17–33 %) for tPA. Conclusions: The tPA with a cutoff of 8.5 ng/mL has a high sensitivity and negative predictive value for exclusion of PE, similar to those observed for the VIDAS D-dimer with a cutoff of 500 ng/mL, although the diagnostic utility was slightly higher for the D-dimer.

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Pestivirus infections in cervids from the Czech Republic

Veterinární Medicína ◽

10.17221/29/2009-vetmed ◽

2009 ◽

Vol 54 (No. 4) ◽

pp. 191-193

Author(s):

K. Sedlak ◽

T. Girma ◽

J. Holejsovsky

Keyword(s):

Czech Republic ◽

Cervus Elaphus ◽

Red Deer ◽

Competitive Inhibition ◽

Enzyme Linked Immunosorbent Assay ◽

Border Disease Virus ◽

Serum Samples ◽

The Czech Republic ◽

Diarrhea Virus ◽

Border Disease

372 sera of cervids from the Czech Republic were examined for antibodies to the bovine viral diarrhea virus (BVDV) and border disease virus (BDV) by competitive-inhibition enzyme-linked immunosorbent assay (ELISA), and for the presence of the BVDV by AgELISA. Antibodies to BVDV/BDV were found in 0.6% (two positive/305 tested) red deer (Cervus elaphus). BVDV/BDV antibodies were not found in four sika deer (Cervus Nippon) and 63 fallow deer (Dama dama). All serum samples were BVDV antigen negative. Our results confirmed that red deer in the Czech Republic are only rarely infected with Pestiviruses. This was the first survey of pestiviruses in farmed and wild cervids in the Czech Republic.

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NESTROFT—A Cost-Effective Mass Screening Tool for the Detection of β-Thalassemia Carrier Status in Anemic Pregnant Women: A Step Toward Reducing the National Disease Burden

Journal of Laboratory Physicians ◽

10.1055/s-0041-1732493 ◽

2021 ◽

Author(s):

Manasi Gosavi ◽

Ramesh Chavan ◽

M. B. Bellad

Keyword(s):

Pregnant Women ◽

Effective Mass ◽

Mass Screening ◽

Predictive Value ◽

Screening Tool ◽

Hematological Parameters ◽

False Negative ◽

Cost Effective ◽

Mean Corpuscular Hemoglobin ◽

Carrier Status

Abstract Introduction β-Thalassemias are inherited hemoglobinopathies commonly encountered in practice. With chances of a promising cure being rare, the prevention of births with this disorder should assume priority, especially in low-resource countries. This can be achieved by the implementation of a mass screening program that is reliable and, at the same time, cost-effective. Objectives This study focuses on the utility of Naked Eye Single Tube Red Cell Osmotic Fragility Test (NESTROFT) as a mass screening tool to detect thalassemia carriers. Hematological parameters that may predict carrier status were also evaluated. Materials and Methods Hemoglobin estimation was performed on all consented pregnant women. If the patient was found to have hemoglobin < 11 g/dL, the blood sample was subjected to other routine hematological tests along with peripheral smear examination. NESTROFT was performed using 0.36% saline solution. Confirmation was done using high-performance liquid chromatography (HPLC). Statistical Analysis Data obtained were tabulated using version 21 of the Statistical Package for Social Sciences. Means, standard deviations, and percentages were used to describe the sample. Chi-square test and Students’ “t” test were used to identify differences between the groups. Results Of 441 pregnant women enrolled, 206 were found to be anemic. Nineteen (9.2%) of the anemic pregnant women were detected to be carriers of hemoglobinopathies. Among the hematological parameters, mean red blood cell count and reticulocyte count were higher, while mean corpuscular hemoglobin concentration was lower in carriers. Also, carriers were more likely to present with microcytic hypochromic anemia. NESTROFT showed a sensitivity of 84.21%, specificity of 96.25%, a positive predictive value of 69.56%, and a negative predictive value of 98.36%. A false-positive result was seen in 3.74% of the tests, while a false negative result was seen in 15.78% of the tests. Conclusions NESTROFT (0.36%) can be used as a simple and cost-effective mass screening tool for the detection of carrier status. This should be followed by confirmation using HPLC or hemoglobin electrophoresis.

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Exercise induced reduction in myocardial work predicts significant coronary artery disease in patients with false negative stress echocardiography

European Heart Journal ◽

10.1093/ehjci/ehaa946.0131 ◽

2020 ◽

Vol 41 (Supplement_2) ◽

Author(s):

A.L Chilingaryan ◽

L.G Tunyan ◽

K.G Adamyan ◽

P.H Zelveyan ◽

L.R Tumasyan ◽

...

Keyword(s):

Coronary Artery Disease ◽

Coronary Artery ◽

Predictive Value ◽

Stress Echocardiography ◽

False Negative ◽

Funding Source ◽

Concentric Hypertrophy ◽

Constructive Work ◽

Artery Disease ◽

Isovolumic Relaxation

Abstract Stress echocardiography (SE) is a reliable technique for the diagnosis of coronary artery disease (CAD) with high sensitivity and specificity. However in patients with small left ventricular (LV) cavity caused by marked concentric hypertrophy the sensitivity of SE is low. We assumed that in patients with false negative SE evaluation of global myocardial work (GW) might detect significant CAD. Methods 238 patents with chest pain (98 female, mean age 61±5 years) without history of CAD were referred to SE for CAD confirmation. 94 (39.5%) patients had negative SE and were enrolled in our study for re-examination. Age and gender matched 50 healthy subjects served as controls. GW index (GWI) was obtained from pressure-strain loops composed from speckle tracking analysis indexed to brachial systolic blood pressure. Global constructive work (GCW) as the sum of positive work due to myocardial shortening during systole and negative work due to lengthening during isovolumic relaxation, global wasted work (GWW) as energy loss by myocardial lengthening in systole and shortening in isovolumic relaxation, and GW efficiency (GWE) as the percentage ratio of constructive work to the sum of constructive work and wasted work were measured after submaximal treadmill SE at the heart rate of 100–110 beats per minute (109±11 s after SE) using EchoPac software by blinded experienced echocardiographer. All patients were referred to coronary angiography after re-examination. Results 42 (44.7%) patients had lower GWI values than the lowest limit of GWI value in controls. These patients had significant reduction in GWI, compared with remaining 52 patients in whom GWI did not differ from those of controls (GWI 1897±112 mmHg% vs 2518±243 mmHg%, p<0.01). GCW, GWE and GWW were comparable between patients with or without reduced GWI (GCW 2283±107mmHg% vs 2321±110 mmHg%, p=NS; GWE 96.9±1.1% vs 97.4±1.2%, p=NS; GWW 57±3 mmHg% vs 53±4 mmHg%, p=NS). 28 (66.7%) of 42 patients with GWI reduction and 8 (15.0%) of 52 patients without GWI reduction had at least one vessel significant CAD. GWI had sensitivity, specificity, and accuracy in detection of CAD 78%, 76%, 77% respectively with 67% positive predictive value, and 85% negative predictive value. 29 (80.5%) patients out of 36 with significant CAD had concentric increase in LVMi compared with true negative SE patients (83±6 g/m2 vs 71±4 g/m2, p<0.01). GWI was the predictor of significant CAD (area under the curve 0.793). Conclusion GWI extends diagnostic power of conventional SE in detection of CAD, especially in patients with smaller LV cavity due to concentric hypertrophy when sensitivity of conventional SE is low. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): Committee of Sience at Ministry of Education of Republic of Armenia

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Validity of ultrasound arterial wall vascularization for assessment of vascular inflammation

European Heart Journal ◽

10.1093/ehjci/ehaa946.2403 ◽

2020 ◽

Vol 41 (Supplement_2) ◽

Author(s):

W Sato ◽

Y Kobayashi ◽

M Otaka ◽

M Unuma ◽

T Yamanaka ◽

...

Keyword(s):

Carotid Stenosis ◽

Predictive Value ◽

Arterial Wall ◽

Carotid Arteries ◽

Vascular Inflammation ◽

False Negative ◽

Intraplaque Hemorrhage ◽

Funding Source ◽

Negative Finding ◽

18F Fdg

Abstract Background Vascular inflammation plays a fundamental role in most vascular diseases including atherosclerosis and vasculitis syndrome, in which arterial wall vascularization (AWV) frequently develops. Visualization of AWV is informative in detecting the vascular inflammation but is challenging. A new ultrasound technique (superb micro-vascular imaging [SMI]) allows the detection of extremely low-velocity flows. We examined an availability of SMI for assessment of the instability of atherosclerotic plaques and the activity of Takayasu arteritis (TA). Methods and results The study consists of two independent and consecutive parts A and B, examined in carotid stenosis (A) and TA (B), respectively. In part A, 12 patients with symptomatic severe carotid stenosis (CS group) scheduled for carotid endarterectomy were enrolled. In six of 12 patients, preoperative ultrasonography with SMI showed intraplaque neovascularization at the plaque shoulder. Postoperatively, histopathology confirmed the neovessels at the corresponding sites of visualized AWV. SMI had a sensitivity of 67%, specificity of 90% for detection of AWV in CS group. In SMI analysis, false positive findings were caused by motion artifact and arterial wall calcification, and a false negative finding is attributed by intraplaque hemorrhage. In part B, 10 patients with TA were enrolled. All patients underwent 18F-FDG-PET/CT, and its vascular uptake were compared with AWV detected by SMI. Bilateral common carotid arteries (CCA), internal carotid arteries and common iliac arteries were examined by SMI. Active vascular 18F-FDG uptake (max SUV >2.1) were found at five sites in three patients, which were not significantly correlated with the prevalence of macaroni sign, increase in C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). Of note, SMI revealed AWV at five sites corresponding to uptake of 18F-FDG, with a sensitivity/specificity of 100% and 98%, positive predictive value 71%, and a negative predictive value 100%. Conclusion SMI enables visualization of AWV at vulnerable plaque in CS patients and at 18F-FDG positive sites in TA patients. SMI has potential as a modality to detect the vascular inflammation. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Grant-in-Aid for Scientific Research, Japan

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Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽

10.1159/000517003 ◽

2021 ◽

pp. 1-6

Author(s):

Salman Khan ◽

Syed Asad Ali Shah ◽

Syed Muhammad Jamal

Keyword(s):

Polymerase Chain Reaction ◽

Viral Genome ◽

Foot And Mouth Disease ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Chain Reaction ◽

Kappa Value ◽

Polymerase Chain ◽

Pcr Assays ◽

Level Of Agreement

Background: Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. Methods: A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. Results: S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (p = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. Conclusions: The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

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PEDV Infection Generates Conformation-Specific Antibodies That Can Be Effectively Detected by a Cell-Based ELISA

Viruses ◽

10.3390/v13020303 ◽

2021 ◽

Vol 13 (2) ◽

pp. 303

Author(s):

Wei-Ting Hsu ◽

Chia-Yu Chang ◽

Chih-Hsuan Tsai ◽

Sung-Chan Wei ◽

Huei-Ru Lo ◽

...

Keyword(s):

Recombinant Baculovirus ◽

Immunocytochemical Staining ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Perfect Agreement ◽

Specific Antibodies ◽

Diarrhea Virus ◽

Serological Studies ◽

A Cell ◽

Infected Insect

Porcine epidemic diarrhea virus (PEDV) is a coronavirus that causes serious and highly contagious enteric disease in swine worldwide. In this study, we constructed a recombinant baculovirus (S-Bac) expressing full-length spike protein of the virulent epidemic genotype 2b (G2b) PEDV strain for serological studies of infected pigs. We found that most spike-specific antibodies produced upon PEDV infection in pigs are conformation-specific and they could be detected on S-Bac-infected insect cells by immunofluorescent assay, but they were insensitive to Western blot analysis, the typical method for antiserum analysis. These results indicated that spike conformation is crucial for serum recognition. Since it is difficult to purify trimeric spike membrane protein for conventional enzyme-linked immunosorbent assay (ELISA), we used S-Bac to generate a novel cell-based ELISA for convenient PEDV detection. We analyzed 100 pig serum samples, and our cell-based ELISA exhibited a sensitivity of 100%, a specificity of 97%, and almost perfect agreement [Cohen’s kappa coefficient value (κ) = 0.98] with immunocytochemical staining results. Our cell-based ELISA rapidly presented antigen for proper detection of conformation-specific antibodies, making PEDV detection more convenient, and it will be useful for detecting many viral diseases in the future.

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Evaluation of the Carba NP Test for Rapid Detection of Carbapenemase-Producing Enterobacteriaceae and Pseudomonas aeruginosa

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00878-13 ◽

2013 ◽

Vol 57 (9) ◽

pp. 4578-4580 ◽

Cited By ~ 147

Author(s):

Nathalie Tijet ◽

David Boyd ◽

Samir N. Patel ◽

Michael R. Mulvey ◽

Roberto G. Melano

Keyword(s):

Pseudomonas Aeruginosa ◽

Positive Predictive Value ◽

Negative Predictive Value ◽

Predictive Value ◽

Rapid Detection ◽

False Negative ◽

Bacterial Extract ◽

Content Type ◽

Negative Results ◽

False Negative Results

ABSTRACTThe Carba NP test was evaluated against a panel of 244 carbapenemase- and non-carbapenemase-producingEnterobacteriaceaeandPseudomonas aeruginosaisolates. We confirmed the 100% specificity and positive predictive value of the test, but the sensitivity and negative predictive value were 72.5% and 69.2%, respectively, and increased to 80% and 77.3%, respectively, using a more concentrated bacterial extract. False-negative results were associated with mucoid strains or linked to enzymes with low carbapenemase activity, particularly OXA-48-like, which has emerged globally in enterobacteria.

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