Prevalence estimation of Tropheryma whipplei in duodenal biopsy tissues of Koreans.

Whipple's disease caused by Tropheryma whipplei is difficult to diagnose because of a broad spectrum of manifestations and non-specific clinical signs. In the current global era, the incidence of duodenal infection/inflammation caused by T. whipplei in Korea may has been underestimated. Here we estimated the prevalence of T. whipplei in duodenal biopsy tissues of Koreans using real-time PCRs (RT-PCRs). A total of 252 duodenal biopsy tissues were collected from Korean patients who underwent esophagogastroduodenoscopy and duodenal biopsy. DNA extracted from the duodenal biopsy tissues was analyzed using three RT-PCRs targeting T. whipplei-specific regions of the 16S-23S rRNA intergenic spacer, hsp65, and Dig15 in parallel. In the samples positive in RT-PCRs, direct sequencing was performed for each RT-PCR target. The prevalence of T. whipplei was estimated based on the RT-PCR and sequencing results. Among the analyzed samples, T. whipplei was not detected. The prevalence of T. whipplei in duodenal biopsy tissues of Koreans was estimated to be less than 0.4%. This is the first study to attempt to detect T. whipplei in duodenal biopsy tissues of Koreans and estimate its prevalence. Our findings infer that while T. whipplei carriers exist in Korea, the incidence of duodenal infection/inflammation caused by T. whipplei is extremely rare.

Performance of digital morphology analyzer CellaVision DC-1.

OBJECTIVES: CellaVision DC-1 (DC-1, Sysmex, Kobe, Japan) is a newly launched digital morphology analyzer that was developed mainly for small to medium-volume laboratories. We evaluated the precision, qualitative performance, comparison of cell counts between DC-1 and manual counting, and turnaround time (TAT) of DC-1. METHODS: Using five peripheral blood smear (PBS) slides spanning normal white blood cell (WBC) range, precision and qualitative performance of DC-1 were evaluated according to the Clinical and Laboratory Standards Institute (CLSI) EP15-A3, EP15-Ed3-IG1, and EP12-A2 guidelines. Cell counts of DC-1 and manual counting were compared according to the CLSI EP 09C-ED3 guidelines, and TAT of DC-1 was also compared with TAT of manual counting. RESULTS: DC-1 showed excellent precision (%CV, 0.0-3.5%), high specificity (98.9-100.0%), and high negative predictive value (98.4-100.0%) in 18 cell classes (12 WBC classes and six non-WBC classes). However, DC-1 showed 0% of positive predictive value in seven cell classes (metamyelocytes, myelocytes, promyelocytes, blasts, plasma cells, nucleated red blood cells, and unidentified). The largest absolute mean differences (%) of DC-1 vs. manual counting was 2.74. Total TAT (min:s) was comparable between DC-1 (8:55) and manual counting (8:55). CONCLUSIONS: This is the first study that comprehensively evaluated the performance of DC-1 including its TAT. DC-1 has a reliable performance that can be used in small to medium-volume laboratories for assisting PBS review. However, DC-1 may make unnecessary workload for cell verification in some cell classes.

Performance of digital morphology analyzer Medica EasyCell assistant.

OBJECTIVES: The EasyCell assistant (Medica, Bedford, MA, USA) is one of the state-of-the-art digital morphology analyzers. We explored the performance of EasyCell assistant in comparison with manual microscopic review and Pentra DX Nexus (Horiba ABX Diagnostics, Montpellier, France). METHODS: In a total of 225 samples (100 normal and 125 abnormal samples), white blood cell (WBC) differentials and platelet (PLT) count estimation by EasyCell assistant were compared with the results by manual microscopic review and Pentra DX Nexus. The manual microscopic review was performed according to the Clinical and Laboratory Standards Institute guidelines (H20-A2). RESULTS: WBC differentials between pre-classification by EasyCell assistant and manual counting showed moderate correlations for neutrophils (r=0.58), lymphocytes (r=0.69), and eosinophils (r=0.51) in all samples. After user verification, they showed mostly high to very high correlations for neutrophils (r=0.74), lymphocytes (r=0.78), eosinophils (r=0.88), and other cells (r=0.91). PLT count by EasyCell assistant highly correlated with that by Pentra DX Nexus (r=0.82). CONCLUSIONS: The performance of EasyCell assistant for WBC differentials and PLT count seems to be acceptable even in abnormal samples with improvement after user verification. The EasyCell assistant, with its reliable performance on WBC differentials and PLT count, would help optimize the workflow of hematology laboratories with reduced workload of manual microscopic review.

Prevalence and detection of Tropheryma whipplei in the stools of Korean patients with diarrhea using real-time PCRs.

BACKGROUND: The prevalence of Tropheryma whipplei varies depending on age, region, and underlying disease. We estimated the prevalence of T. whipplei in the stools of Korean patients with diarrhea using real-time PCR (RT-PCR) and compared three RT-PCR targets, rpoB, hsp65, and Dig15. METHODS: A total of 1404 nucleic acid samples extracted from the stools of Korean patients with diarrhea were tested using an initial RT-PCR targeting T. whipplei-specific regions of 16S-23S rRNA intergenic spacer. Subsequently, the samples positive for the initial RT-PCR were tested using the follow-up RT-PCRs targeting rpoB, hsp65, and Dig15 and analyzed by sequencing to confirm the presence of T. whipplei. We estimated the prevalence of T. whipplei and compared them according to gender and age. We also compared the performance of three targets in the follow-up RT-PCRs. RESULTS: T. whipplei was detected in 1.4% of all samples (20 of 1404), and there were no differences according to gender and age. In pediatric samples (≤ 19 years), T. whipplei was detected higher in children aged 6-19 than in those aged 1-5 (2.7% vs. 0.7%, P = 0.01). Sensitivities of the rpoB, hsp65, and Dig15 RT-PCR were 50.0%, 85.0%, and 95.0%, respectively; specificities were 100.0%, 100.0%, and 84.6%, respectively. CONCLUSIONS: This is the first study that estimated the prevalence of T. whipplei in the stools of Korean patients with diarrhea. This study demonstrated the presence of T. whipplei in stools of Koreans, even though the bacterium was detected low. The RT-PCRs targeting hsp65 and Dig15 showed reliable performance, and a multiplex PCR including these targets is expected to be useful for T. whipplei detection.

Performance of digital morphology analyzer Vision Pro on white blood cell differentials.

OBJECTIVES: Vision Pro (West Medica, Perchtoldsdorf, Austria) is a recently developed digital morphology analyzer. We evaluated the performance of Vision Pro on white blood cell (WBC) differentials. METHODS: In a total of 200 peripheral blood smear samples (100 normal and 100 abnormal samples), WBC preclassification and reclassification by Vision Pro were evaluated and compared with manual WBC count, according to the Clinical and Laboratory Standards Institute guidelines (H20-A2). RESULTS: The overall sensitivity was high for normal WBCs and nRBCs (80.1-98.0%). The overall specificity and overall efficiency were high for all cell classes (98.1-100.0% and 97.7-99.9%, respectively). The absolute values of mean differences between Vision Pro and manual count ranged from 0.01 to 1.31. In leukopenic samples, those values ranged from 0.09 to 2.01. For normal WBCs, Vision Pro preclassification and manual count showed moderate or high correlations (r=0.52-0.88) except for basophils (r=0.34); after reclassification, the correlation between Vision Pro and manual count was improved (r=0.36-0.90). CONCLUSIONS: This is the first study that evaluated the performance of Vision Pro on WBC differentials. Vision Pro showed reliable analytical performance on WBC differentials with improvement after reclassification. Vision Pro could help improve laboratory workflow.

Evaluation of high-throughput digital lateral flow immunoassays for the detection of influenza A/B viruses from clinical swab samples.

We evaluated the performance of new high-throughput digital lateral flow immunoassays (LFIAs) detecting influenza antigens and compared them with those of the widely used digital LFIA and the rapid nucleic acid amplification test (NAAT). We tested 199 clinical nasopharyngeal (nasal) swab samples using three LFIA tests (BD Veritor Plus, STANDARD F Influenza A/B FIA, and ichroma TRIAS) and the rapid NAAT (ID NOW Influenza A & B2). Agreements and clinical performances (sensitivity and specificity) were evaluated based on the results of reverse transcriptase-polymerase chain reaction (RT-PCR) and verification panel. The agreement of each test with RT-PCR was moderate to almost perfect. The sensitivity of ID NOW was significantly higher than that of LFIAs (P = .0005, .0044, and .0026 for influenza A and P = .0044, .0026, and .0044 for influenza B, respectively). The specificities were not significantly different between the four tests (P > .05). However, the reference panel suggests that ichroma TRIAS test is more sensitive than the other two LFIA tests. All three LFIA assays performed similarly with no false positives against influenza A. For influenza B, ichroma TRIAS had 2 of 166 false positives whereas there were no false positives for the other two LFIA tests. Influenza antigen digital LFIAs have advantages in terms of the workflow when simultaneous tests are required. Rapid NAAT has higher sensitivity, while new antigen LFIAs are efficient and high-throughput. It is recommended that users select appropriate methods and algorithms according to the number of specimens and laboratory conditions in each clinical laboratory.

Performance evaluation of microfluidic microplate-based fluorescent ELISA for qualitative detection of SARS-CoV-2-specific IgG and IgM.

We evaluated the diagnostic performance of newly developed microfluidic microplate-based fluorescent ELISA for anti-SARS-CoV-2 antibody detection: the Veri-Q opti COVID-19 IgG and IgM ELISAs (hereafter, "Opti IgG/M"; MiCo BioMed, Gyeonggi-do, Republic of Korea), in comparison with conventional ELISAs. A total of 270 serum samples were analyzed, among which 90 samples were serially obtained from 25 COVID-19 patients. Another 180 samples were collected from 180 SARS-CoV-2-negative individuals. As comparative assays, we used SCoV-2 Detect IgG/M ELISA (hereafter, "InBios IgG/M"; InBios, Seattle, WA, USA) and Veri-Q COVID-19 IgG/IgM ELISA (hereafter, "Veri-Q IgG/M"; MiCo BioMed). Compared with conventional ELISAs, the Opti IgG yielded 97.1-100.0% positive percent agreement, 95.2-98.0% negative percent agreement, 96.3-97.8% total percent agreement, and kappa values of 0.90-0.94. Between the Opti IgM and the InBios IgM, the values were 93.7%, 96.6%, 95.9%, and 0.89, respectively. For the Opti IgG, sensitivities for the samples collected from 0-7, 8-14, 15-21, and ≥ 22 days after symptom onset were 40.0, 58.3, 94.1, and 100.0%, respectively. The values for the Opti IgM were 30.0, 54.2, 88.2, and 80%, respectively. The diagnostic specificities of the Opti IgG and IgM were 99.4 and 97.2%, respectively. The microfluidic microplate-based fluorescent ELISAs showed comparable diagnostic performance to conventional ELISAs for detecting anti-SARS-CoV-2 antibodies. With the combination of high throughput, a simplified workflow, and the ability to analyze reduced volumes, this new technology has great potential for improving SARS-CoV-2 serologic testing.

Analytical performance of the digital morphology analyzer Sysmex DI-60 for body fluid cell differential counts.

BACKGROUND: Sysmex DI-60 (Sysmex, Kobe, Japan) is a digital morphology (DM) analyzer widely used in clinical laboratories and supports body fluid (BF) applications. We evaluated analytical performance of DI-60 compared with XN-350 (Sysmex) and manual counting for BF cell differential counts. METHODS: A total of 213 BF samples were collected (47 cerebrospinal fluid [CSF], 80 pleural fluid, and 86 ascites samples). The analytical performance of DI-60 for BF cell differential counts was evaluated based on sensitivity, specificity, accuracy, and agreement. BF cell differential counts obtained by DI-60 were compared with those obtained by XN-350 and manual counting. RESULTS: The overall sensitivity was high for neutrophils, lymphocytes, and macrophages (range, 83.1-99.4%). The overall specificity and overall accuracy were high for all cell types (range, 95.3-99.7% and 94.3-99.3%, respectively). The agreement between DI-60 pre-classification and verification was strong (κ = 0.89). The absolute mean differences between DI-60 verification and XN-350 ranged from 0.26 to 11.05, and differences between DI-60 verification and manual counting ranged from 0.01 to 4.76. CONCLUSIONS: This is the first study to evaluate the performance of DI-60 compared with XN-350 and manual counting for BF cell differential counts. DI-60 showed reliable performance with CSF, pleural fluid, and ascites samples. For BF cell differential counts, DI-60 may be a better option than XN-350 and could be used for screening purposes in understaffed laboratories. To improve the hematology workflow for BF cell differential counting, the DM analyzer needs to be optimized by taking into account the laboratory situation and unmet needs, and the clinical laboratory needs to establish criteria for verification and manual slide review.

Clinical performance of SARS-CoV-2 antigen-detection rapid diagnostic test using SERS-based lateral flow immunoassay.

Background: 'ACROSIS COVID-19 Ag (NPS)' kit (SG Medical, Seoul, Korea) is a newly developed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen-detection rapid diagnostic test (Ag-RDT) using surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFIA). We evaluated its clinical performance compared with STANDARD Q COVID-19 Ag (SD Biosensor, Suwon, Korea), a previously approved Ag-RDT. Methods: A total of 286 nasopharyngeal swab specimens were collected: 104 positive and 182 negative specimens in SARS-CoV-2 real-time reverse-transcription polymerase-chain-reaction (rRT-PCR). SARS-CoV-2-positive specimens were divided according to the cycle threshold (Ct) value in rRT-PCR. The clinical performance of ACROSIS was compared with that of STANDARD Q. Results: ACROSIS showed significantly higher sensitivity than STANDARD Q (92.3% vs. 85.6%, P = 0.02), especially in specimens with 25 ≤ Ct < 30 (78.6% vs. 42.9%). The Ct values of RdRp/S genes for 95% detection rates by ACROSIS and STANDARD Q were 25.8 and 23.0, respectively. Conclusions: This is the first study that evaluated the performance of ACROSIS compared with STANDARD Q. The overall clinical performance of ACROSIS was superior to that of STANDARD Q, especially in specimens with 25 ≤ Ct < 30. ACROSIS could be useful for SARS-CoV-2 Ag detection even in relatively low viral load specimens.

Exploring the vaccine-induced immunity against severe acute respiratory syndrome coronavirus 2 in healthcare workers.

We aimed to analyze the kinetics of T-cell-mediated and B-cell-mediated humoral immune responses against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) before and after booster vaccination, as well as the impacts of the in vitro test results the type of vaccination on the prediction of SARS-CoV-2 infection. A total of 240 healthcare workers vaccinated twice were serially tested using an interferon gamma release assay (IGRA) and a neutralizing antibody (nAb). At the end of the study, we investigated the history of SARS-CoV-2 infection of all the enrolled participants to analyze the effects of the test results and the type of vaccination on SARS-CoV-2 infection. Overall, the positive rates were 52.3% and 80.0% for IGRA and 84.6% and 100% for the nAb test before and after booster vaccination, respectively. However, the positive rates were 52.8% for IGRA and 100% for nAb 3 months after booster vaccination. The in vitro test results and the type of vaccination were not associated with SARS-CoV-2 infection. The antibody response caused by the SARS-CoV-2 vaccination lasted more than 6 months, although the response of the T-cells disappeared rapidly after 3 months. However, these in vitro results and the type of vaccination cannot be used for predicting the risk of SARS-CoV-2 infection.

Establishment of Reference Intervals of Cytokeratin 19 Fragment Antigen 21-1 in Korean Adults.

Cytokeratin 19 fragment antigen 21-1 (CYFRA 21-1) is useful for predicting and monitoring non-small cell lung cancer prognosis. We established reference intervals (RIs) of CYFRA 21-1 in Korean adults, including those older than 60 years. Data of 4,098 apparently healthy subjects (age range, 20-87 years) were analyzed after excluding those with a history of malignancy, high tumor marker concentrations (except CYFRA 21-1), and/or abnormal findings on a chest computed tomography scan through medical chart review. After removing two outliers, RIs of CYFRA 21-1 were determined using data of 4,096 subjects based on the non-parametric method (2.5th and 97.5th percentiles) according to CLSI guidelines EP28-A3c. The subjects were divided into two and four groups according to sex and age (20-40, 41-50, 51-60, and >60 years), respectively, and the median CYFRA 21-1 concentration was compared between the groups. The RI of CYFRA 21-1 was 0.66-3.84 ng/mL, applicable to both men and women. Regardless of sex, the CYFRA 21-1 concentration increased with age, suggesting that age-dependent RIs of CYFRA 21-1 should be applied. Rather than using a single RI provided by the manufacturer, the RI of CYFRA 21-1 should be continually verified and established in each clinical laboratory.

Clinical Impact of Recipient-Derived Isoagglutinin Levels in ABO-Incompatible Hematopoietic Stem Cell Transplantation.

ABO incompatibility is not considered a contraindication for hematopoietic stem cell transplantation (HSCT). We hypothesized that recipient-derived isoagglutinin (RDI) levels could play a critical role in clinical outcomes. In this study, we compared clinical outcomes such as survival, GVHD, infection, relapse, transfusion, and engraftment, among ABO-compatible patients (ABOc), ABO-incompatible patients (ABOi) with low RDI, and ABOi patients with high RDI. The ABOi with high RDI group was defined as recipients with more than 1:16 RDI levels. We analyzed 103 recipients (ABOc, 53; ABOi with low RDI, 36; ABOi with high RDI, 14). The ABOi with high RDI group showed a decreased 1-year survival and increased acute GVHD grade IV and RBC transfusion (p = 0.017, 0.027, and 0.032, respectively). The ABOi with high RDI group was an independent risk factor for increased death, RBC transfusion, and poor platelet (PLT) engraftment (odds ratio (OR) = 3.20, p = 0.01; OR = 8.28, p = 0.02; OR = 0.18, p = 0.03, respectively). The ABOi with high RDI group showed significantly delayed PLT engraftment. In conclusion, this is the first study underscoring high RDI levels as a marker predicting unfavorable outcomes in ABOi HSCT.

Reference Interval for the Axis-Shield Clinical Chemistry Heparin-Binding Protein Assay.

The newly developed Axis-Shield clinical chemistry heparin-binding protein (HBP) assay (Axis-Shield Diagnostics Ltd., Dundee, Scotland) can be applied to fully automated platforms. We aimed to establish a reference interval (RI) of HBP using the Axis-Shield HBP assay, and to evaluate the analytical performance of this assay. An RI was established in 212 sodium citrated plasma samples using the non-parametric method (2.5th and 97.5th percentiles). Precision, linearity, and carry-over were evaluated according to the Clinical and Laboratory Standards Institute guidelines. The RI of HBP was between 5.3 ng/mL and 171.0 ng/mL, which could be applied regardless of gender and age. Percentage coefficients of variations (%CVs) of repeatability and within-laboratory precision were 4.9% and 6.3%, respectively, for low-concentration control and 1.6% and 3.0%, respectively, for high-concentration control. The linearity was excellent (coefficient of determination (R2) = 0.99), and the carry-over rate was negligible (0.05%). This is the first study to establish an RI of HBP using the newly developed and fully automated Axis-Shield HBP assay. The Axis-Shield HBP assay showed an acceptable level of analytical performance and could be used to measure HBP concentrations effectively in routine clinical practice. Further studies are awaited to evaluate the clinical utility of HBP using this automated assay.

Digital Morphology Analyzer Sysmex DI-60 vs. Manual Counting for White Blood Cell Differentials in Leukopenic Samples: A Comparative Assessment of Risk and Turnaround Time.

BACKGROUND: Digital morphology (DM) analyzers are increasingly being used for white blood cell (WBC) differentials. We assessed the laboratory efficiency of the Sysmex DI-60 system (DI-60; Sysmex, Kobe, Japan) in comparison with manual counting in leukopenic samples. METHODS: In total, 40 peripheral blood smear samples were divided into normal, mild leukopenia, moderate leukopenia, and severe leukopenia groups based on WBC count. In each group, the risk and turnaround time (TAT) were compared between DI-60 and manual counting. Risk was determined by failure mode and effect analysis using the risk priority number (RPN) score, and TAT was recorded for the analytical phase. RESULTS: Overall, DI-60 showed a five-fold lower risk (70 vs. 350 RPN) and longer TAT than manual counting. In severe leukopenic samples, DI-60 showed a shorter TAT/slide and a remarkably lower cell count/slide than manual counting. In all samples, the TAT/cell for DI-60 was substantially longer than that for manual counting (DI-60 vs. manual: total, 1.8 vs. 1.0 sec; normal, 1.5 vs. 0.7 sec; mild leukopenia, 1.9 vs. 0.9 sec; moderate leukopenia, 1.8 vs. 1.0 sec; severe leukopenia, 28.8 vs. 19.0 sec). CONCLUSIONS: This is the first comparative assessment of risk and TAT between DI-60 and manual counting in leukopenic samples. DI-60 decreases the laboratory risk and improves patient safety, but requires more time to count fewer cells, especially in severe leukopenic samples. DM analyzers should be applied selectively depending on the WBC count to optimize laboratory efficiency.

Clinical Performance of Two Automated Immunoassays, EliA CTD Screen and QUANTA Flash CTD Screen Plus, for Antinuclear Antibody Screening.

BACKGROUND: Recently, two fully automated immunoassays for antinuclear antibody (ANA) screening were introduced: EliA CTD Screen (Thermo Fisher Scientific, Freiburg, Germany) and QUANTA Flash CTD Screen Plus (Inova Diagnostics, San Diego, USA). We evaluated their clinical performance in comparison with the indirect immunofluorescence assay (IIFA) and analyzed samples with discrepant results. METHODS: In total, 406 serum samples (206 from patients undergoing routine checkups and 200 from rheumatology clinic patients) were assayed using EliA, QUANTA Flash, and IIFA. We evaluated assay concordance and agreement and confirmed the presence of anti-extractable nuclear antigen (ENA) antibodies in samples with discrepant automated immunoassay and IIFA results. Additionally, we compared the clinical performance of each assay in diagnosing ANA-associated rheumatic disease (AARD) and adjusted the cut-off values. RESULTS: In rheumatology clinic samples, the concordance and agreement were 91.5% and strong between EliA and QUANTA Flash, 79.0% and weak between EliA and IIFA, and 80.5% and moderate between QUANTA Flash and IIFA, respectively. In automated immunoassay-positive, IIFA-negative samples (N=15), all anti-ENA antibodies detected (6/15) were anti-Sjögren's syndrome antigen A/Ro (Ro60) antibodies. The automated immunoassays and IIFA showed high accuracy for diagnosing AARD, and adjusted cut-off values improved their sensitivities (EliA with 0.56 ratio, 82.9% sensitivity; QUANTA Flash with 9.7 chemiluminescent units, 87.8% sensitivity). CONCLUSIONS: The two automated immunoassays showed reliable performance compared with IIFA and can be efficiently used with the IIFA in clinical immunology laboratories. Clinical cut-off values can be adjusted according to the workflow in each laboratory.

How Reproducible Is the Data from Sysmex DI-60 in Leukopenic Samples?

Digital morphology (DM) analyzers are widely applied in clinical practice. It is necessary to evaluate performances of DM analyzers by focusing on leukopenic samples. We evaluated the analytical performance, including precision, of a Sysmex DI-60 system (Sysmex, Kobe, Japan) on white blood cell (WBC) differentials in leukopenic samples. In a total of 40 peripheral blood smears divided into four groups according to WBC count (normal, mild, moderate, and severe leukopenia; each group n = 10), we evaluated precision of WBC preclassificaiton by DI-60. %coefficients of variation (%CVs) of precision varied for each sample and for each cell class; the fewer cells per slide, the higher %CV. The overall specificity and efficiency were high for all cell classes except plasma cells (95.9-99.9% and 90.0-99.4%, respectively). The largest absolute value of mean difference between DI-60 and manual count in each group was: 10.77, normal; 10.22, mild leukopenia; 19.09, moderate leukopenia; 47.74, severe leukopenia. This is the first study that evaluated the analytical performance of DI-60 on WBC differentials in leukopenic samples as the main subject. DI-60 showed significantly different performance depending on WBC count. DM analyzers should be evaluated separately in leukopenic samples, even if the overall performance was acceptable.

Performance evaluation of coaguchek pro II in comparison with coaguchek XS plus and sta-r Max using a sta-neoplastine CI plus.

INTRODUCTION: We evaluated the analytical performance of CoaguChek Pro II (Roche Diagnostics GmbH, Mannheim, Germany), a new point-of-care device measuring the international normalized ratio (INR) values, in comparison with CoaguChek XS Plus (Roche Diagnostics GmbH) and STA-R Max using STA-Neoplastine CI Plus (Diagnostica Stago SAS, Asnières-sur-Seine, France). METHODS: The precision of Pro II was analyzed, according to the Clinical and Laboratory Standards Institute guidelines (CLSI POCT14-A2 and EP15-A3). In 105 clinical samples, the Pro II INR values were compared with those of XS Plus and STA-R Max using STA-Neoplastine CI Plus (CLSI EP09-A3 and EP35). We also compared the Pro II INR values between capillary blood (CB) and venous blood (VB; CLSI EP35). RESULTS: The precision of Pro II was acceptable (within-run and between-run CV%: 2.71% and 3.28% at normal level; 1.52% and 4.47% at abnormal level, respectively). The Pro II INR values showed very high correlation and almost perfect agreement with those of XS Plus and STA-R Max using STA-Neoplastine CI Plus (r = .97 and κ = .94; r = .95 and κ = .91). The mean difference between Pro II and STA-R Max using STA-Neoplastine CI Plus increased as INR values increased, with 60% of samples showing differences >0.5 in the supratherapeutic range. The Pro II INR values showed very high correlation between CB and VB (r = .98). CONCLUSION: Pro II INR values are accurate and reliable using both CB and VB; however, they should be confirmed by laboratory analyzers in the supratherapeutic range.

Performance Evaluation of the QXDx BCR-ABL %IS Droplet Digital PCR Assay.

Accurate detection of BCR-ABL fusion transcripts at and below molecular response (MR) 4 (0.01% International Scale [IS]) is required for disease monitoring in patients with chronic myeloid leukemia (CML). We evaluated the analytical performance of the QXDx BCR-ABL %IS (Bio-Rad, Hercules, CA, USA) droplet digital PCR (ddPCR) assay, which is the first commercially available ddPCR-based in vitro diagnostics product. In precision analysis, the %CV was 9.3% and 3.0%, with mean values of 0.031% IS and 9.4% IS, respectively. The assay was linear in the first order, ranging from 0.032% IS to 20% IS. The manufacturer-claimed limit of blank, limit of detection, and limit of quantification were verified successfully. There was a very strong correlation between the results of the QXDx BCR-ABL %IS ddPCR assay and the ipsogen BCR-ABL1 Mbcr IS-MMR (Qiagen, Hilden, Germany) real-time quantitative PCR assay (r=0.996). In conclusion, the QXDx BCR-ABL %IS ddPCR assay can provide reliable results for CML patients.