Interferon-gamma release assays (IGRAs) are widely used to identify latent tuberculosis infection (LTBI); however, inherent test variability affects their diagnostic interpretation. We evaluated false-positive and false-negative rates, as well as conversion and reversion rates across CVs of 20%, 40%, 60%, 80%, and 100%, using statistical modeling. At a diagnostic cutoff of 0.35 IU/mL, false-negative rates increased from 1.61% to 33.41% with increasing CVs, whereas false-positive rates ranged from 0.00% to 15.87% within the 0.20-0.70 IU/mL borderline range. Expanding the borderline to 0.20-1.00 IU/mL reduced false-positive rates to a maximum of 3.16%, without affecting false-negative rates. Within the 0.20-0.70 IU/mL borderline zone, correct reversion and false conversion rates at 0.20 and 0.35 IU/mL ranged from 0.01% to 25.00% and 0.00% to 24.20%, respectively. At 0.35, 0.70, and 1.00 IU/mL, correct conversion and false reversion rates ranged from 0.05% to 24.20% and 0.00% to 25.00%, respectively. These results highlight the importance of adopting borderline zones in IGRA interpretation to reduce misclassification, although variability from manufacturing, pre-analytical processing, and analytical procedures remains a significant challenge. Reducing such variability through improved production consistency, standardized sample handling, and automated analysis platforms is essential to enhance the diagnostic reliability of IGRAs for LTBI.
{"title":"Effect of Variability on Interferon-Gamma Release Assay Performance: A Quantitative Analysis.","authors":"Won-Ki Min","doi":"10.3343/alm.2025.0066","DOIUrl":"https://doi.org/10.3343/alm.2025.0066","url":null,"abstract":"Interferon-gamma release assays (IGRAs) are widely used to identify latent tuberculosis infection (LTBI); however, inherent test variability affects their diagnostic interpretation. We evaluated false-positive and false-negative rates, as well as conversion and reversion rates across CVs of 20%, 40%, 60%, 80%, and 100%, using statistical modeling. At a diagnostic cutoff of 0.35 IU/mL, false-negative rates increased from 1.61% to 33.41% with increasing CVs, whereas false-positive rates ranged from 0.00% to 15.87% within the 0.20-0.70 IU/mL borderline range. Expanding the borderline to 0.20-1.00 IU/mL reduced false-positive rates to a maximum of 3.16%, without affecting false-negative rates. Within the 0.20-0.70 IU/mL borderline zone, correct reversion and false conversion rates at 0.20 and 0.35 IU/mL ranged from 0.01% to 25.00% and 0.00% to 24.20%, respectively. At 0.35, 0.70, and 1.00 IU/mL, correct conversion and false reversion rates ranged from 0.05% to 24.20% and 0.00% to 25.00%, respectively. These results highlight the importance of adopting borderline zones in IGRA interpretation to reduce misclassification, although variability from manufacturing, pre-analytical processing, and analytical procedures remains a significant challenge. Reducing such variability through improved production consistency, standardized sample handling, and automated analysis platforms is essential to enhance the diagnostic reliability of IGRAs for LTBI.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"14 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144693366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BackgroundCancer cell line-derived feeder cells enhance natural killer (NK) cell expansion; however, concerns regarding viable residual feeder cells in the final product limit their use. Evidence supporting the safety of NK-sensitive K562-based feeders, even when irradiated, is scarce. We optimized an NK cell expansion protocol using genetically engineered K562-mbIL-18/-21 (GE-K562) feeder cells and clinical-grade media and confirmed the absence of residual feeder cells.MethodsNK cell expansion efficiency was compared between feeder-free and feeder-based systems using CTS NK-Xpander Medium. To achieve optimal NK expansion, various peripheral blood mononuclear cell (PBMC)-to-feeder ratios and re-stimulation frequencies were tested over 21 days. Flow cytometry and BCR::ABL1 quantitative reverse transcription PCR (RT-qPCR) were used to confirm the absence of feeder cells in the final NK cell product.ResultsFeeder-based systems showed superior NK cell fold expansion compared with that of feeder-free systems. Among feeder-based conditions, NK cells expanded 5,224-fold at a 2:1 PBMC-to-feeder ratio after 3 weeks, relative to 1,450-fold at a 6:1 ratio (P <0.05). Re-stimulation on days 7 and 14 further increased expansion up to 261,457-fold. Irradiated feeder cells showed no proliferation and were eliminated within 3-6 days. On day 21, flow cytometry and BCR::ABL1 RT-qPCR results confirmed the absence of residual feeder cells.ConclusionsOur optimized NK cell expansion protocol using irradiated GE-K562 feeder cells and clinical-grade media offers a safe and scalable approach to generating large numbers of NK cells, supporting its potential use in clinical immunotherapy applications.
肿瘤细胞系来源的饲养细胞增强自然杀伤细胞（NK）的扩增；然而，对最终产品中残留的饲养细胞的担忧限制了它们的使用。支持对nk敏感的基于k562的喂食器的安全性的证据，即使经过辐照，也很少。我们使用基因工程K562-mbIL-18/-21 （GE-K562）饲养细胞和临床级培养基优化了NK细胞扩增方案，并证实没有残留的饲养细胞。方法采用CTS nk - expander培养基，比较无投料系统和有投料系统的snk细胞扩增效率。为了达到最佳NK扩增，在21天内测试了不同的外周血单核细胞（PBMC）与饲料的比例和再刺激频率。用流式细胞术和BCR::ABL1定量反转录PCR （RT-qPCR）证实最终NK细胞产物中不存在饲养细胞。结果加料系统的NK细胞折叠扩增率高于无加料系统。在以饲料为基础的条件下，在2:1的pbmc与饲料比例下，NK细胞在3周后扩增5,224倍，而在6:1的比例下扩增1,450倍（P <0.05）。第7天和第14天的再次刺激进一步增加了膨胀，达到261,457倍。辐照后的饲养细胞无增殖，3-6天内消失。第21天，流式细胞术和BCR::ABL1 RT-qPCR结果证实没有残留的饲养细胞。结论采用辐照的GE-K562饲养细胞和临床级培养基进行NK细胞扩增，是一种安全、可扩展的大量NK细胞扩增方法，支持其在临床免疫治疗中的潜在应用。
{"title":"Optimization of Natural Killer Cell Expansion with K562-mbIL-18/-21 Feeder Cells and Assurance of Feeder Cell-free Products.","authors":"Hantae Jo,Yujung Jo,Seung Kwon Koh,Jinho Kim,SoonHo Kweon,Jeehun Park,Hyun-Young Kim,Duck Cho,Mijeong Lee","doi":"10.3343/alm.2025.0168","DOIUrl":"https://doi.org/10.3343/alm.2025.0168","url":null,"abstract":"BackgroundCancer cell line-derived feeder cells enhance natural killer (NK) cell expansion; however, concerns regarding viable residual feeder cells in the final product limit their use. Evidence supporting the safety of NK-sensitive K562-based feeders, even when irradiated, is scarce. We optimized an NK cell expansion protocol using genetically engineered K562-mbIL-18/-21 (GE-K562) feeder cells and clinical-grade media and confirmed the absence of residual feeder cells.MethodsNK cell expansion efficiency was compared between feeder-free and feeder-based systems using CTS NK-Xpander Medium. To achieve optimal NK expansion, various peripheral blood mononuclear cell (PBMC)-to-feeder ratios and re-stimulation frequencies were tested over 21 days. Flow cytometry and BCR::ABL1 quantitative reverse transcription PCR (RT-qPCR) were used to confirm the absence of feeder cells in the final NK cell product.ResultsFeeder-based systems showed superior NK cell fold expansion compared with that of feeder-free systems. Among feeder-based conditions, NK cells expanded 5,224-fold at a 2:1 PBMC-to-feeder ratio after 3 weeks, relative to 1,450-fold at a 6:1 ratio (P <0.05). Re-stimulation on days 7 and 14 further increased expansion up to 261,457-fold. Irradiated feeder cells showed no proliferation and were eliminated within 3-6 days. On day 21, flow cytometry and BCR::ABL1 RT-qPCR results confirmed the absence of residual feeder cells.ConclusionsOur optimized NK cell expansion protocol using irradiated GE-K562 feeder cells and clinical-grade media offers a safe and scalable approach to generating large numbers of NK cells, supporting its potential use in clinical immunotherapy applications.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"32 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144693310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In Young Yoo,Sungjin Jo,Joo An Kwon,Jay Ho Han,Hae Kyung Lee,Yeon-Joon Park
BackgroundVarious molecular methods are used to rapidly detect gastrointestinal pathogens, highlighting the importance of understanding the performance of the associated kits in detail. We comprehensively assessed the performance of Kogene PowerChek multiplex real-time PCR kits (PowerChek Bacterial/Viral Kits) with the FilmArray GI Panel.MethodsResidual stool specimens (N=246), initially tested utilizing the FilmArray GI Panel (May 2023-Jan 2024), were reanalyzed using PowerChek Bacterial/Viral Kits. Discrepancies were resolved by performing additional molecular assays and reviewing culture results when available. True positives (TPs)/true negatives were defined by concordant results in at least two assays. We determined cycle threshold (Ct)/crossing point (Cp) distributions between the TP and false positive (FP) groups and analyzed melting curves for the FilmArray GI Panel FPs.ResultsThe positive-percent agreement (PPA) of the PowerChek Bacterial/Viral Kits was 50-100%, with lower values for Salmonella spp., rotavirus, and astrovirus, whereas the FilmArray GI Panel showed 100% PPA across all targets. Both platforms demonstrated >99% negative-percent agreement, except for enteropathogenic Escherichia coli (EPEC) and adenovirus (PowerChek Bacterial/Viral Kits) or EPEC, enteroaggregative E. coli, norovirus, and Salmonella spp. (FilmArray GI Panel). The FPs showed higher Ct/Cp values with both kits, and these values were significantly higher for adenovirus (PowerChek Viral Kit), EPEC, and norovirus (FilmArray GI Panel). Melting curve analysis of four norovirus FPs (FilmArray GI Panel) revealed atypical patterns in three cases.ConclusionsThe FilmArray GI Panel demonstrated higher sensitivity than the PowerChek Kits. For norovirus, melting curve analysis will help avoid FPs.
各种分子方法用于快速检测胃肠道病原体，强调了详细了解相关试剂盒性能的重要性。我们使用FilmArray GI Panel综合评估了Kogene PowerChek多重实时PCR试剂盒（PowerChek细菌/病毒试剂盒）的性能。方法使用FilmArray GI Panel（2023年5月- 2024年1月）进行初步检测的残余粪便标本（N=246），使用PowerChek细菌/病毒试剂盒重新分析。差异通过进行额外的分子分析和检查培养结果来解决。真阳性(TPs)/真阴性由至少两次试验的一致结果定义。我们确定了TP组和假阳性组之间的循环阈值(Ct)/交叉点（Cp）分布，并分析了FilmArray GI Panel FPs的熔化曲线。结果powercheck细菌/病毒试剂盒的阳性率（PPA）为50-100%，对沙门氏菌、轮状病毒和星状病毒的阳性率较低，而FilmArray GI Panel对所有靶标的阳性率为100%。除了肠致病性大肠杆菌（EPEC）和腺病毒（powercheck细菌/病毒试剂盒）或EPEC、肠聚集性大肠杆菌、诺如病毒和沙门氏菌（FilmArray GI Panel）外，这两个平台均显示出99%阴性的一致性。两种试剂盒的FPs均显示较高的Ct/Cp值，并且腺病毒（powercheck病毒试剂盒）、EPEC和诺如病毒（FilmArray GI Panel）的这些值显著较高。4例诺如病毒FPs的熔化曲线分析（FilmArray GI Panel）显示3例不典型。结论FilmArray GI Panel比PowerChek Kits具有更高的灵敏度。对于诺如病毒，熔化曲线分析有助于避免FPs。
{"title":"Kogene PowerChek Multiplex Real-time PCR Kits Versus the BioFire FilmArray Gastrointestinal Panel: Roles of Crossing Point Values and Melting Curves in Interpreting the FilmArray Gastrointestinal Panel.","authors":"In Young Yoo,Sungjin Jo,Joo An Kwon,Jay Ho Han,Hae Kyung Lee,Yeon-Joon Park","doi":"10.3343/alm.2025.0047","DOIUrl":"https://doi.org/10.3343/alm.2025.0047","url":null,"abstract":"BackgroundVarious molecular methods are used to rapidly detect gastrointestinal pathogens, highlighting the importance of understanding the performance of the associated kits in detail. We comprehensively assessed the performance of Kogene PowerChek multiplex real-time PCR kits (PowerChek Bacterial/Viral Kits) with the FilmArray GI Panel.MethodsResidual stool specimens (N=246), initially tested utilizing the FilmArray GI Panel (May 2023-Jan 2024), were reanalyzed using PowerChek Bacterial/Viral Kits. Discrepancies were resolved by performing additional molecular assays and reviewing culture results when available. True positives (TPs)/true negatives were defined by concordant results in at least two assays. We determined cycle threshold (Ct)/crossing point (Cp) distributions between the TP and false positive (FP) groups and analyzed melting curves for the FilmArray GI Panel FPs.ResultsThe positive-percent agreement (PPA) of the PowerChek Bacterial/Viral Kits was 50-100%, with lower values for Salmonella spp., rotavirus, and astrovirus, whereas the FilmArray GI Panel showed 100% PPA across all targets. Both platforms demonstrated >99% negative-percent agreement, except for enteropathogenic Escherichia coli (EPEC) and adenovirus (PowerChek Bacterial/Viral Kits) or EPEC, enteroaggregative E. coli, norovirus, and Salmonella spp. (FilmArray GI Panel). The FPs showed higher Ct/Cp values with both kits, and these values were significantly higher for adenovirus (PowerChek Viral Kit), EPEC, and norovirus (FilmArray GI Panel). Melting curve analysis of four norovirus FPs (FilmArray GI Panel) revealed atypical patterns in three cases.ConclusionsThe FilmArray GI Panel demonstrated higher sensitivity than the PowerChek Kits. For norovirus, melting curve analysis will help avoid FPs.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"661 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144669338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BackgroundSingle nucleotide polymorphism-based chromosomal microarray analysis (CMA) can detect regions of homozygosity (ROHs), which may be associated with medical conditions; however, limited ROH data, especially in East Asians, complicates clinical interpretations. We characterized ROH distributions and frequencies in a Korean population using CMA, highlighting clinically relevant findings, including suspected uniparental disomy (UPD), using standardized criteria.MethodsWe analyzed ROHs in 1,731 individuals who underwent postnatal CMA at a Korean medical center. ROHs ≥ 3 Mb long were detected using the CytoScan Dx platform and Chromosome Analysis Suite Dx. Suspected UPD and consanguinity were assessed per the American College of Medical Genetics and Genomics technical standards.ResultsWe identified 3,962 ROHs, with 76.7% of patients carrying at least one. Common "hotspot" regions included 3p21.31p21.1 (20.3%), 11p11.2 (18.2%), 1q21.1q21.3 (17.7%), and 1p33p32.3 (12.0%). Almost all ROHs observed in >1% of patients had a median size of <5 Mb. ROH frequencies correlated negatively with chromosomal recombination rates and positively with gene densities. Additionally, 1.2% (N = 21) of patients exhibited ROH patterns suggestive of UPD or consanguinity (13 suspected UPDs on imprinted chromosomes, 6 on non-imprinted chromosomes, and 2 consanguinities); 8 of 13 patients with suspected UPD were diagnosed as having imprinting disorders, with no pathogenic copy number variations detected.ConclusionsOur population-specific ROH data for Koreans improve clinical interpretations by minimizing the risk of overinterpreting benign variants and highlight the value of standardized criteria for reliably detecting UPD and consanguinity and integrating ROH analysis into routine CMA interpretations.
{"title":"Regions of Homozygosity Identified with a Chromosomal Microarray in a Korean Population: Distribution, Frequency, and Clinical Interpretation.","authors":"Jaeryuk Kim,Sunghee Min,Chang Ahn Seol,Eul-Ju Seo","doi":"10.3343/alm.2025.0021","DOIUrl":"https://doi.org/10.3343/alm.2025.0021","url":null,"abstract":"BackgroundSingle nucleotide polymorphism-based chromosomal microarray analysis (CMA) can detect regions of homozygosity (ROHs), which may be associated with medical conditions; however, limited ROH data, especially in East Asians, complicates clinical interpretations. We characterized ROH distributions and frequencies in a Korean population using CMA, highlighting clinically relevant findings, including suspected uniparental disomy (UPD), using standardized criteria.MethodsWe analyzed ROHs in 1,731 individuals who underwent postnatal CMA at a Korean medical center. ROHs ≥ 3 Mb long were detected using the CytoScan Dx platform and Chromosome Analysis Suite Dx. Suspected UPD and consanguinity were assessed per the American College of Medical Genetics and Genomics technical standards.ResultsWe identified 3,962 ROHs, with 76.7% of patients carrying at least one. Common \"hotspot\" regions included 3p21.31p21.1 (20.3%), 11p11.2 (18.2%), 1q21.1q21.3 (17.7%), and 1p33p32.3 (12.0%). Almost all ROHs observed in >1% of patients had a median size of <5 Mb. ROH frequencies correlated negatively with chromosomal recombination rates and positively with gene densities. Additionally, 1.2% (N = 21) of patients exhibited ROH patterns suggestive of UPD or consanguinity (13 suspected UPDs on imprinted chromosomes, 6 on non-imprinted chromosomes, and 2 consanguinities); 8 of 13 patients with suspected UPD were diagnosed as having imprinting disorders, with no pathogenic copy number variations detected.ConclusionsOur population-specific ROH data for Koreans improve clinical interpretations by minimizing the risk of overinterpreting benign variants and highlight the value of standardized criteria for reliably detecting UPD and consanguinity and integrating ROH analysis into routine CMA interpretations.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"24 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144652888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BackgroundAn international reference measurement laboratory network for creatinine (Cr) is lacking; therefore, Korea developed an independent evaluation and certification system. The in vitro diagnostics (IVD) certification program, launched in 2017, formed part of a broader Cr standardization initiative intended to enhance accuracy at the manufacturing stage.MethodsThe program was designed to evaluate analytical systems, including all reagent lots, calibrators, and instrument models, twice annually. Bias, imprecision, total error (TE), and linearity were evaluated based on established acceptance criteria. A post-certification process allows submission for a second challenge and validation of corrective actions.ResultsBetween 2017 and 2023, 489 analytical systems were evaluated. Average acceptance rates for bias, imprecision, TE, and linearity were 70.8%, 95.9%, 87.7%, and 87.8%, respectively. The lowest acceptance rate for bias evaluation was 8.7% for the kinetic Jaffe method without compensation in 2018. Over the 7-year period, the mean absolute percentage bias (absBias%), coefficient of variation (CV), and TE were 4.62%, 1.37%, and 7.29%, respectively. The highest absBias% (7.94%) was observed in the 0.0 ≤ Cr < 1.0 target value range. Since 2019, a consistent reduction in absBias% has been observed.ConclusionsThis program is a pioneering response to the absence of a global certification program for Cr assays. It offers significant advantages, including comprehensive evaluations, fee-free participation, and a robust post-certification process. Continuous participation and improvement efforts by manufacturers have contributed to enhanced accuracy in Cr assays.
{"title":"In Vitro Diagnostics Certification for Creatinine Assays in Korea over 7 Years: Achievements and Future Outlook.","authors":"Eun-Jung Cho,Joonsang Yu,Jeayeon Ryu,Jiwoo Seo,Hyunae Lee,Chan-Ik Cho,Tae-Dong Jeong,Sollip Kim,Woochang Lee,Sail Chun,Won-Ki Min","doi":"10.3343/alm.2024.0654","DOIUrl":"https://doi.org/10.3343/alm.2024.0654","url":null,"abstract":"BackgroundAn international reference measurement laboratory network for creatinine (Cr) is lacking; therefore, Korea developed an independent evaluation and certification system. The in vitro diagnostics (IVD) certification program, launched in 2017, formed part of a broader Cr standardization initiative intended to enhance accuracy at the manufacturing stage.MethodsThe program was designed to evaluate analytical systems, including all reagent lots, calibrators, and instrument models, twice annually. Bias, imprecision, total error (TE), and linearity were evaluated based on established acceptance criteria. A post-certification process allows submission for a second challenge and validation of corrective actions.ResultsBetween 2017 and 2023, 489 analytical systems were evaluated. Average acceptance rates for bias, imprecision, TE, and linearity were 70.8%, 95.9%, 87.7%, and 87.8%, respectively. The lowest acceptance rate for bias evaluation was 8.7% for the kinetic Jaffe method without compensation in 2018. Over the 7-year period, the mean absolute percentage bias (absBias%), coefficient of variation (CV), and TE were 4.62%, 1.37%, and 7.29%, respectively. The highest absBias% (7.94%) was observed in the 0.0 ≤ Cr < 1.0 target value range. Since 2019, a consistent reduction in absBias% has been observed.ConclusionsThis program is a pioneering response to the absence of a global certification program for Cr assays. It offers significant advantages, including comprehensive evaluations, fee-free participation, and a robust post-certification process. Continuous participation and improvement efforts by manufacturers have contributed to enhanced accuracy in Cr assays.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"44 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144639839","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Taehwa Kim,Daesup Lee,Woo Hyun Cho,Sun Min Lee,Kyung-Hwa Shin,Hye Ju Yeo
BackgroundThe utility of the QuantiFERON-Monitor (QFM, Qiagen), a tool developed to assess general immune function, remains insufficiently explored in critically ill patients with acute respiratory failure (ARF). Therefore, we used the QFM to evaluate the immune function of patients with ARF at intensive care unit (ICU) admission and monitored QFM changes based on disease severity and clinical outcome correlations.MethodsWe evaluated the immune function of 99 patients with ARF in an ICU setting. The QFM was evaluated upon ICU admission, day 7 post-ICU admission, and discharge. Their results were compared with those of five healthy controls.ResultsThe QFM levels at ICU admission were significantly lower in patients with ARF than in healthy controls (median IUs/mL: 5.5 vs. 465.0, respectively). The QFM levels in patients with coronavirus disease 2019 or pneumonia (9.2 and 7.9 IUs/mL, respectively) were higher than those in patients with acute respiratory distress syndrome or septic shock (4.9 and 3.6 IUs/mL, respectively). On day 7, the QFM levels increased to 8.3 IUs/mL and reached 16.7 IUs/mL at discharge. At ICU admission, patients requiring ventilator support had lower QFM levels than those requiring nasal prong or high-flow nasal cannula support. Those who died in the ICU had significantly lower QFM levels (4.0 IUs/mL) at ICU admission than those who survived (5.8 IUs/mL).ConclusionsReduced QFM levels among patients with severe ARF reflect impaired cellular immune responses and suggest that QFM may serve as a practical tool for early risk stratification and immune monitoring in ICU settings.
QuantiFERON-Monitor （QFM, Qiagen）是一种用于评估一般免疫功能的工具，但在急性呼吸衰竭（ARF）危重患者中的应用仍未得到充分探索。因此，我们使用QFM来评估ARF患者在重症监护病房（ICU）入院时的免疫功能，并根据疾病严重程度和临床结局相关性监测QFM的变化。方法对ICU收治的99例ARF患者进行免疫功能评估。在ICU入院、ICU入院后第7天和出院时评估QFM。他们的结果与五名健康对照者的结果进行了比较。结果ARF患者入院时的QFM水平明显低于健康对照组（中位iu /mL: 5.5 vs 465.0）。2019冠状病毒病和肺炎患者的QFM水平（分别为9.2和7.9 IUs/mL）高于急性呼吸窘迫综合征和感染性休克患者（分别为4.9和3.6 IUs/mL）。第7天，QFM水平上升至8.3 IUs/mL，放电时达到16.7 IUs/mL。在ICU入院时，需要呼吸机支持的患者的QFM水平低于需要鼻尖或高流量鼻插管支持的患者。ICU死亡患者入院时QFM水平（4.0 iu /mL）明显低于存活患者（5.8 iu /mL）。结论严重急性肾功能衰竭患者QFM水平降低反映了细胞免疫反应受损，提示QFM可作为ICU环境中早期风险分层和免疫监测的实用工具。
{"title":"Role of the QuantiFERON-Monitor in Assessing the Immune Status of Patients with Acute Respiratory Failure in Adult Intensive Care Units: A Prospective, Observational Study.","authors":"Taehwa Kim,Daesup Lee,Woo Hyun Cho,Sun Min Lee,Kyung-Hwa Shin,Hye Ju Yeo","doi":"10.3343/alm.2024.0691","DOIUrl":"https://doi.org/10.3343/alm.2024.0691","url":null,"abstract":"BackgroundThe utility of the QuantiFERON-Monitor (QFM, Qiagen), a tool developed to assess general immune function, remains insufficiently explored in critically ill patients with acute respiratory failure (ARF). Therefore, we used the QFM to evaluate the immune function of patients with ARF at intensive care unit (ICU) admission and monitored QFM changes based on disease severity and clinical outcome correlations.MethodsWe evaluated the immune function of 99 patients with ARF in an ICU setting. The QFM was evaluated upon ICU admission, day 7 post-ICU admission, and discharge. Their results were compared with those of five healthy controls.ResultsThe QFM levels at ICU admission were significantly lower in patients with ARF than in healthy controls (median IUs/mL: 5.5 vs. 465.0, respectively). The QFM levels in patients with coronavirus disease 2019 or pneumonia (9.2 and 7.9 IUs/mL, respectively) were higher than those in patients with acute respiratory distress syndrome or septic shock (4.9 and 3.6 IUs/mL, respectively). On day 7, the QFM levels increased to 8.3 IUs/mL and reached 16.7 IUs/mL at discharge. At ICU admission, patients requiring ventilator support had lower QFM levels than those requiring nasal prong or high-flow nasal cannula support. Those who died in the ICU had significantly lower QFM levels (4.0 IUs/mL) at ICU admission than those who survived (5.8 IUs/mL).ConclusionsReduced QFM levels among patients with severe ARF reflect impaired cellular immune responses and suggest that QFM may serve as a practical tool for early risk stratification and immune monitoring in ICU settings.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"266 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144594226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Youngjae Huh,Jaebon Lee,Inha Hwang,Ye Eun Yoon,Eun Jin Lee,Taekyu Lim,Jae Won Yun
Although splenomegaly is typically uncommon in myelodysplastic syndromes (MDS), it is associated with reduced engraftment rates and poor survival outcomes. Despite its clinical significance, the incidence and genetic associations of splenomegaly in MDS remain understudied. To address this, we conducted a retrospective study of 27 patients with MDS at the Veterans Health Service Medical Center in South Korea. Based on computed tomography scan evaluation, splenomegaly was identified in 26% of patients with MDS, and significant associations with variants in ASXL1 (P=0.0089 for null and missense/inframe variants) and RUNX1 (P=0.042 for null variants) were observed, suggesting that these variants are linked to an increased risk of splenomegaly. Notably, one patient with ASXL1 and TET2 variants developed severe splenomegaly (spleen size, 29 cm) following granulocyte colony-stimulating factor (G-CSF) treatment, requiring splenectomy. This case suggests a potential interaction between specific genetic variants and G-CSF sensitivity, potentially exacerbating splenomegaly. Our findings suggest that the incidence of splenomegaly in patients with MDS, including mild cases, is likely underestimated and that ASXL1 and RUNX1 variants increase the risk of splenomegaly. Furthermore, careful monitoring for the development of severe splenomegaly during G-CSF treatment may be warranted in genetically susceptible individuals with MDS.
{"title":"Association of ASXL1 and RUNX1 Variants with Splenomegaly in MDS Based on Next-generation Sequencing and Computed Tomography Data: A Retrospective Study.","authors":"Youngjae Huh,Jaebon Lee,Inha Hwang,Ye Eun Yoon,Eun Jin Lee,Taekyu Lim,Jae Won Yun","doi":"10.3343/alm.2025.0033","DOIUrl":"https://doi.org/10.3343/alm.2025.0033","url":null,"abstract":"Although splenomegaly is typically uncommon in myelodysplastic syndromes (MDS), it is associated with reduced engraftment rates and poor survival outcomes. Despite its clinical significance, the incidence and genetic associations of splenomegaly in MDS remain understudied. To address this, we conducted a retrospective study of 27 patients with MDS at the Veterans Health Service Medical Center in South Korea. Based on computed tomography scan evaluation, splenomegaly was identified in 26% of patients with MDS, and significant associations with variants in ASXL1 (P=0.0089 for null and missense/inframe variants) and RUNX1 (P=0.042 for null variants) were observed, suggesting that these variants are linked to an increased risk of splenomegaly. Notably, one patient with ASXL1 and TET2 variants developed severe splenomegaly (spleen size, 29 cm) following granulocyte colony-stimulating factor (G-CSF) treatment, requiring splenectomy. This case suggests a potential interaction between specific genetic variants and G-CSF sensitivity, potentially exacerbating splenomegaly. Our findings suggest that the incidence of splenomegaly in patients with MDS, including mild cases, is likely underestimated and that ASXL1 and RUNX1 variants increase the risk of splenomegaly. Furthermore, careful monitoring for the development of severe splenomegaly during G-CSF treatment may be warranted in genetically susceptible individuals with MDS.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"8 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144568619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BackgroundIron deficiency (ID) and hemoglobinopathies are highly prevalent in Southeast Asia. Accurate estimation of reference intervals (RIs) for red cell parameters is complicated by the need to exclude individuals with these conditions from the reference population. Indirect RI estimations using machine learning could help overcome these challenges.MethodsWe developed a binary classification model using eXtreme Gradient Boosting (XGB) to distinguish normal individuals from those with ID, hemoglobinopathies, or other anemias. The model was trained on an annotated dataset comprising 5,520 complete blood count (CBC) results and validated with a holdout dataset of 2,367 CBC results. An independent dataset of 64,100 CBC results was used to identify individuals predicted to be normal, from which RIs were estimated using the refineR algorithm.ResultsThe XGB model achieved an area under the ROC of 0.97 (95% confidence interval: 0.96-0.97) for distinguishing between individuals with normal versus abnormal values. Among individuals within the independent dataset, 40,300 (62.9%) were predicted to be normal. The refineR-based reference limits (RLs) derived from this subset approximated those obtained through a direct approach. Improvements in the accuracy of indirect RL estimates were most evident for hematocrit, hemoglobin, and red cell concentrations.ConclusionsCombining XGB with refineR to indirectly derive RIs for red cell parameters improved the accuracy and yielded results comparable with those of directly derived RIs. A further benefit was the capacity to generate sex- and age-specific ranges, which has remained difficult to achieve through direct approaches.
{"title":"A Machine Learning Approach to Reference Interval Estimation for Red Cell Parameters in a South and East Asian Population.","authors":"Veera Sekaran Nadarajan,Pavai Sthaneshwar,Jia Qi Lim,Angeli Ambayya,Putri Junaidah Megat Yunus","doi":"10.3343/alm.2025.0027","DOIUrl":"https://doi.org/10.3343/alm.2025.0027","url":null,"abstract":"BackgroundIron deficiency (ID) and hemoglobinopathies are highly prevalent in Southeast Asia. Accurate estimation of reference intervals (RIs) for red cell parameters is complicated by the need to exclude individuals with these conditions from the reference population. Indirect RI estimations using machine learning could help overcome these challenges.MethodsWe developed a binary classification model using eXtreme Gradient Boosting (XGB) to distinguish normal individuals from those with ID, hemoglobinopathies, or other anemias. The model was trained on an annotated dataset comprising 5,520 complete blood count (CBC) results and validated with a holdout dataset of 2,367 CBC results. An independent dataset of 64,100 CBC results was used to identify individuals predicted to be normal, from which RIs were estimated using the refineR algorithm.ResultsThe XGB model achieved an area under the ROC of 0.97 (95% confidence interval: 0.96-0.97) for distinguishing between individuals with normal versus abnormal values. Among individuals within the independent dataset, 40,300 (62.9%) were predicted to be normal. The refineR-based reference limits (RLs) derived from this subset approximated those obtained through a direct approach. Improvements in the accuracy of indirect RL estimates were most evident for hematocrit, hemoglobin, and red cell concentrations.ConclusionsCombining XGB with refineR to indirectly derive RIs for red cell parameters improved the accuracy and yielded results comparable with those of directly derived RIs. A further benefit was the capacity to generate sex- and age-specific ranges, which has remained difficult to achieve through direct approaches.","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":"92 1","pages":""},"PeriodicalIF":4.9,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144547854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Exemestane, an aromatase inhibitor commonly used for breast cancer treatment, shares structural similarities with sex steroids analyzed in clinical laboratories. We aimed to investigate the influence of exemestane cross-reactivity in the measurement of sex steroids across various immunoassays.
Methods: We conducted a multicenter study involving measurements of androstenedione, testosterone, estradiol, progesterone, and 17-hydroxyprogesterone in serum samples from women undergoing exemestane therapy (N=15; 25 mg/day). Measurements were performed using liquid chromatography-mass spectrometry (LC-MS) and various commercially available chemiluminescence immunoassays, ELISA, and radioimmunoassay. In-vitro cross-reactivity was assessed by adding exemestane and 17-hydroexemestane to serum samples.
Results: Patients undergoing exemestane therapy had markedly falsely elevated androstenedione results in all immunoassays evaluated (N=4), which correlated with serum exemestane levels. In-vitro experiments confirmed this interference to be caused by cross-reactivity with exemestane. Additionally, one immunoassay yielded falsely elevated estradiol results in 20% of patients. However, in-vitro experiments did not confirm this to be caused by cross-reactivity with exemestane or 17-hydroexemestane.
Conclusions: Exemestane cross-reacts with androstenedione immunoassays, causing falsely elevated results in treated patients. This analytical interference may raise unnecessary concerns, leading to expensive diagnostic workups.
{"title":"Analytical Interference of Exemestane With Androstenedione Immunoassays.","authors":"Marina Giralt, Roser Ferrer, Noelia Díaz-Troyano, Belén Vega, Manuel Luque-Ramírez, Sílvia Martínez, Bárbara Fernández, Irene Martínez, Aleix Fabregat, Eulalia Urgell, Ignacio Cardona, Gregori Casals, Héctor F Escobar-Morreale","doi":"10.3343/alm.2024.0362","DOIUrl":"10.3343/alm.2024.0362","url":null,"abstract":"<p><strong>Background: </strong>Exemestane, an aromatase inhibitor commonly used for breast cancer treatment, shares structural similarities with sex steroids analyzed in clinical laboratories. We aimed to investigate the influence of exemestane cross-reactivity in the measurement of sex steroids across various immunoassays.</p><p><strong>Methods: </strong>We conducted a multicenter study involving measurements of androstenedione, testosterone, estradiol, progesterone, and 17-hydroxyprogesterone in serum samples from women undergoing exemestane therapy (N=15; 25 mg/day). Measurements were performed using liquid chromatography-mass spectrometry (LC-MS) and various commercially available chemiluminescence immunoassays, ELISA, and radioimmunoassay. In-vitro cross-reactivity was assessed by adding exemestane and 17-hydroexemestane to serum samples.</p><p><strong>Results: </strong>Patients undergoing exemestane therapy had markedly falsely elevated androstenedione results in all immunoassays evaluated (N=4), which correlated with serum exemestane levels. <i>In-vitro</i> experiments confirmed this interference to be caused by cross-reactivity with exemestane. Additionally, one immunoassay yielded falsely elevated estradiol results in 20% of patients. However, <i>in-vitro</i> experiments did not confirm this to be caused by cross-reactivity with exemestane or 17-hydroexemestane.</p><p><strong>Conclusions: </strong>Exemestane cross-reacts with androstenedione immunoassays, causing falsely elevated results in treated patients. This analytical interference may raise unnecessary concerns, leading to expensive diagnostic workups.</p>","PeriodicalId":8421,"journal":{"name":"Annals of Laboratory Medicine","volume":" ","pages":"410-419"},"PeriodicalIF":4.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12187492/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143668880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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