scholarly journals Analytical Performance of the Sysmex HISCL HBsAg Assay and Comparison with the Roche Elecsys HBsAg II Quant Assay in the Quantification of Hepatitis B Surface Antigen

Medicina ◽  
2021 ◽  
Vol 57 (12) ◽  
pp. 1307
Author(s):  
Joonhong Park ◽  
Taewon Bae ◽  
Yonggon Cho ◽  
Dalsik Kim ◽  
Jaehyeon Lee
Keyword(s):  
Comparative Evaluation ◽  
Hepatitis B Surface Antigen ◽  
Reference Value ◽  
Analytical Performance ◽  
Clinical Samples ◽  
Limit Of Quantification ◽  
Highly Sensitive ◽  
Hbsag Assay ◽  
Allowable Error ◽  
Assay Precision

Background and Objectives: This study aims to estimate the analytical performance of the Sysmex HISCL HBsAg assay and to assess the analytical correlation with the Roche Elecsys HBsAg II quant assay with clinical samples and the WHO International Standard (IS). Materials and Methods: The intra-assay precision, linearity, assay limitation, accuracy, and comparative evaluation of the HISCL HBsAg assay were estimated. Results: Extrapolating from the plot of the average total allowable error versus the reference value, an accuracy goal of 20% would be achieved around a limit of quantification (LoQ) of 0.014867 IU/mL. The percentage of biases for each level of the WHO IS measured by the two assays were less than 15%, except for the WHO 3rd IS, for which the HISCL HBsAg assay achieved a percentage of bias of 33%. In the comparative evaluation, Passing–Bablok regression analysis did not reveal any significant deviation from linearity between the two assays (y = −48.6998 + 1.9206x; p = 0.79 by the CUSUM test for linearity). The mean difference of the quantitative HBsAg level between the two assays was 1762.5 IU/mL in the Bland–Altman plot. Conclusions: The HISCL HBsAg assay, with a highly sensitive LoQ of 0.03 IU/mL, showed similar analytical performance in HBsAg quantification to the Elecsys HBsAg II quant assay and may be helpful in obtaining better diagnoses and therapeutic strategies for treating HBV infections.

scholarly journals Analytical performance and method comparison of a quantitative point-of-care immunoassay for measurement of bile acids in cats and dogs

2020 ◽  
Vol 33 (1) ◽  
pp. 35-46
Author(s):  
Kristina Weiler ◽  
Katharina Kleber ◽  
Sabine Zielinsky ◽  
Andreas Moritz ◽  
Natali Bauer
Keyword(s):  
Bile Acids ◽  
Point Of Care ◽  
Method Comparison ◽  
Reference Method ◽  
Analytical Performance ◽  
Total Serum ◽  
Limit Of Quantification ◽  
Coefficients Of Variation ◽  
Test Kit ◽  
Allowable Error

Point-of-care analyzers (POCAs) for quantitative assessment of bile acids (BAs) are scarce in veterinary medicine. We evaluated the Fuji Dri-Chem Immuno AU10V analyzer and v-BA test kit (Fujifilm) for detection of feline and canine total serum BA concentration. Results were compared with a 5th-generation assay as reference method and a 3rd-generation assay, both run on a bench-top analyzer. Analytical performance was assessed at 3 different concentration ranges, and with interferences. For method comparison, samples of 60 healthy and diseased cats and 64 dogs were included. Linearity was demonstrated for a BA concentration up to 130 µmol/L in cats ( r = 0.99) and 110 µmol/L in dogs ( r = 0.99). The analyzer showed high precision near the lower limit of quantification of 2 µmol/L reported by the manufacturer. Intra- and inter-assay coefficients of variation were < 5% for both species and all concentrations. Interferences were observed for bilirubin (800 mg/L) and lipid (4 g/L). There was excellent correlation with the reference method for feline ( rs = 0.98) and canine samples ( rs = 0.97), with proportional biases of 6.7% and −1.3%, respectively. However, a large bias (44.1%) was noted when the POCA was compared to the 3rd-generation assay. Total observed error was less than total allowable error at the 3 concentrations. The POCA reliably detected feline and canine BA in clinically relevant concentrations.

scholarly journals Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Qrt Pcr ◽  
Highly Sensitive ◽  
One Step ◽  
Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

scholarly journals Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chukwunonso Onyilagha ◽  
Henna Mistry ◽  
Peter Marszal ◽  
Mathieu Pinette ◽  
Darwyn Kobasa ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Real Time ◽  
Point Of Care ◽  
Middle Eastern ◽  
Turnaround Time ◽  
Cross Reactivity ◽  
Clinical Samples ◽  
Diarrhea Virus ◽  
Highly Sensitive ◽  
Transmissible Gastroenteritis

AbstractThe coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.

scholarly journals HCV Detection, Discrimination, and Genotyping Technologies

Sensors ◽  
2018 ◽  
Vol 18 (10) ◽  
pp. 3423 ◽  
Author(s):  
Shrikant Warkad ◽  
Satish Nimse ◽  
Keum-Soo Song ◽  
Taisun Kim
Keyword(s):  
Hepatitis C ◽  
Hcv Infection ◽  
World Health ◽  
Clinical Samples ◽  
Infected People ◽  
Advantages And Disadvantages ◽  
Highly Sensitive ◽  
The World ◽  
Health Organization

According to the World Health Organization (WHO), 71 million people were living with Hepatitis C virus (HCV) infection worldwide in 2015. Each year, about 399,000 HCV-infected people succumb to cirrhosis, hepatocellular carcinoma, and liver failure. Therefore, screening of HCV infection with simple, rapid, but highly sensitive and specific methods can help to curb the global burden on HCV healthcare. Apart from the determination of viral load/viral clearance, the identification of specific HCV genotype is also critical for successful treatment of hepatitis C. This critical review focuses on the technologies used for the detection, discrimination, and genotyping of HCV in clinical samples. This article also focuses on advantages and disadvantages of the reported methods used for HCV detection, quantification, and genotyping.

scholarly journals Total Fat Gravimetric Method Workflow in Portuguese Olives Using Closed-Vessel Microwave-Assisted Extraction (MAE)

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2364
Author(s):  
João Gonçalo Lourenço ◽  
Daniel Ettlin ◽  
Inês Carrero Cardoso ◽  
Jesus M. Rodilla
Keyword(s):  
Limit Of Detection ◽  
Gravimetric Method ◽  
Reference Value ◽  
Microwave Assisted Extraction ◽  
Closed Vessel ◽  
Limit Of Quantification ◽  
Microwave Assisted ◽  
Assisted Extraction ◽  
Total Fat

A simple and rapid method for the quantitation of total fat in olive samples is designed, evaluated, and presented. This method is based on an innovative closed-vessel microwave-assisted extraction (MAE) technique. A method was designed for olives, and some figures of merits were evaluated: limit of detection (LOD), limit of quantification (LOQ) and expanded uncertainty (U). The data obtained in these experiences show that the workflow of the MAE method in a closed container is statistically equivalent to the other two methods, showing in this case better performance indicators (LOD = 0.02%, LOQ = 0.06%, and U = 15%). In addition, it is also demonstrated that the complete MAE method workflow allows the determination of total fat in a maximum of 12 analyses simultaneously for about 100 min in each run, which is the capacity of the rotor. This is a much better productivity when compared to the traditional Soxhlet-based method. Considering the sample workflow, the closed-vessel MAE method greatly simplifies sample handling, therefore minimizing sample loss during sample preparation and reducing analysis time. When MAE is compared to NIR-based methods, the advantage comes from there being no need for any type of calibration in the sample matrix. The MAE method itself can be used to determine the reference value for NIR calibration purposes. The results obtained for CRM using MAE were equivalent to the ones shown on the certificate.

Application of the CLSI EP15-A3 Guideline as an Alternative Troubleshooting Tool for Verification of Assay Precision

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S88-S88
Author(s):  
Jose Jara Aguirre ◽  
Karl Ness ◽  
Alicia Algeciras-Schimnich
Keyword(s):  
Quality Control ◽  
Carcinoembryonic Antigen ◽  
Laboratory Investigation ◽  
Analysis Of Variance ◽  
Experimental Approach ◽  
Biological Variation ◽  
Analytical Performance ◽  
Microsoft Excel ◽  
Before And After ◽  
Assay Precision

Abstract Introduction The CLSI EP15-A3 guideline “User Verification of Precision and Estimation of Bias” provides a simple experimental approach to estimate a method’s imprecision and bias. The objective is to determine if the laboratory precision performance of repeatability (SR) and within-laboratory imprecision (SWL) are in accordance to the manufacturer specification claims (MSCs). Objectives Evaluate the utility of the EP15-A3 protocol to verify method precision during a troubleshooting investigation and after major instrument maintenance, using a carcinoembryonic antigen (CEA) immunoassay as an example. Methods CEA was performed on the Beckman Coulter DxI (Beckman Coulter, Brea, CA). Quality control (QC) levels (L1: 2.89; L2: 21.10; L3: 39.10 ng/mL) (Bio-Rad Laboratories, Irvine, CA) were used. Each QC level was measured before and after instrument maintenance as follows: five replicates per run, one run per day, and during 5 days. Imprecision estimates (IEs) for SR (%CVR) and SWL (%CVWL) were calculated by one-way analysis of variance using Microsoft Excel Analyse-it software. Estimated imprecision was compared to MSC and desirable imprecision specifications based on biological variation (BV). Results A change in the analytical performance of CEA was detected by a decreased sigma-metric indicator. After a bias problem was ruled out, the observed %CVR for L1, L2, and L3 were 7.2%, 3.7%, and 4.8%, respectively. The %CVWL were 8.3%, 5.0%, and 5.5%, which exceeded the MSC of %CVWL~4.0% to 4.5%. After a laboratory investigation, major instrument maintenance was performed by the manufacturer. The %CVR and %CVWL estimates for L1, L2, and L3 after maintenance were 3.2%, 3.8%, 3.5% and 3.9%, 4.2%, 4.0%, respectively. After maintenance, the CEA performance was consistent with the MSC for each of the levels analyzed and within the BV impression goal of %CV ≤6.4. Conclusion CLSI EP15-A3 guideline is an alternative troubleshooting tool that can be used to investigate and verify method precision performance before and after significant instrument maintenance.

scholarly journals The Use of Factorial Design and Simplex Optimization to Improve Analytical Performance of In Situ Film Electrodes

Sensors ◽  
2020 ◽  
Vol 20 (14) ◽  
pp. 3921
Author(s):  
Matjaž Finšgar ◽  
Klara Jezernik
Keyword(s):  
Tap Water ◽  
Optimization Procedure ◽  
Fractional Factorial ◽  
Analytical Performance ◽  
Limit Of Quantification ◽  
Simplex Optimization ◽  
Factors Affecting ◽  
Five Factors ◽  
The Individual

This work presents a systematic approach to determining the significance of the individual factors affecting the analytical performance of in-situ film electrode (FE) for the determination of Zn(II), Cd(II), and Pb(II). Analytical parameters were considered simultaneously, where the lowest limit of quantification, the widest linear concentration range, and the highest sensitivity, accuracy, and precision of the method evidenced a better analytical method. Significance was evaluated by means of a fractional factorial (experimental) design using five factors, i.e., the mass concentrations of Bi(III), Sn(II), and Sb(III), to design the in situ FE, the accumulation potential, and the accumulation time. Next, a simplex optimization procedure was employed to determine the optimum conditions for these factors. Such optimization of the in situ FE showed significant improvement in analytical performance compared to the in situ FEs in the initial experiments and compared to pure in situ FEs (bismuth-film, tin-film, and antimony-film electrodes). Moreover, using the optimized in situ FE electrode, a possible interference effect was checked for different species and the applicability of the electrode was demonstrated for a real tap water sample.

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