Volume 48, Issue 4

Liquid biopsy (LB) represents an advanced, minimally invasive approach that elevates the precision of oncological decision-making by identifying tumor DNA in bodily fluids. However, despite numerous endorsements from international specialty societies and working groups, implementation of LB into routine care is lagging behind due to conceptual and methodological uncertainties. This concise mini review aims to help catalyzing the translation of LB into routine care by exploring key considerations for incorporating circulating tumor DNA (ctDNA) analysis into clinical practice. Addressing eight pertinent questions from the perspective of a molecular oncology laboratory, this review synthesizes insights from the European Society for Medical Oncology (ESMO) recommendations and incorporates the latest findings from relevant literature, offering a comprehensive guide to the implementation of ctDNA assays.

Objectives Altered serum Netrin-1 levels have been widely reported in cancer and other clinical diseases and they are often measured by commercial ELISA kits. However, we found the questionable results using these kits and therefore performed this simple study to evaluate their accuracy in detection of serum Netrin-1. Methods Four commonly used commercial kits were collected. The kit standards were serially diluted or spiked into serum samples. The cells with confirmed expression of Netrin-1 and their culture medium, as well as the Netrin-1 controls of each kit were used for the kits to detect. The cell lysate samples and the kit controls were also blotted on a nitrocellulose membrane for detection antibodies of each kit to probe. Results Detection of the Netrin-1 standards in serum by each kit were all affected. Only one kit was able to detect Netrin-1 in the cell lysate or medium. No ELISA kits could detect all Netrin-1 controls of the four kits. None of the detection antibodies correctly probed Netrin-1 in the dot blot. Conclusions The accuracy of these four Netrin-1 ELISA kits is under question. Reported serum Netrin-1 levels based on measurements by these kits need be carefully interpreted.

Objectives The accuracy of blood glucose measurement in clinical laboratories is vital for diabetes diagnosis. Trueness Verification Plan was carried out and analyzed for evaluating the standardization of serum glucose among clinical laboratories. Methods Trueness verification samples were distributed to clinical laboratories for three days measurement, and their target values were assigned by two certified reference laboratories. The relative bias, coefficient of variation (CV), and total error (TE) for each clinical laboratory were calculated and analyzed. Moreover, the Six Sigma metrics and Quality Goal Index were utilized to reflect the measurement quality of the clinical laboratories. Results The pass rates evaluated by bias, CV, and TE ranged from 45.2 % to 64.8 %, 96.8 %–98.9 %, and 83.9 %–97.1 % over the six years. The matched systems used in clinical laboratories demonstrated better accuracy than the un-matched systems. The pass rate by bias of hexokinase method is 53.1 %–78.6 %, while the glucose oxidase method is 29.2 %–52.2 %. Overall, 74.2 %–85.7 % of clinical laboratories achieved an acceptable level (both σ>3), and 35.2 %–61.4 % of laboratories reached a “world-class” level (both σ>6). Conclusions The quality for serum glucose measurement has been greatly improved. However, standardization among clinical systems still needs to be further promoted.

Objectives Biobanks play an important role in advancing cancer research, yet concerns persist regarding the molecular integrity of long-term stored samples. This study assessed fresh frozen (FF) tissues and formalin-fixed paraffin-embedded (FFPE) tissues from the Siriraj Hospital colorectal cancer (CRC) biobank collected during two distinct periods (2011–2012 and 2020–2021). Methods In 2022, FF and FFPE primary cancer tissues from 75 CRC patients were evaluated. RNA sequencing (RNA-Seq) analyzed comprehensive gene expression profiles in FF tissues preserved at −80 °C, while nCounter profiling elucidated cancer-specific RNA transcripts in FFPE tissues stored at ambient temperature. Comparative analyses were conducted between specimens from 2011 to 2012 and 2020–2021. Results The FF tissues stored for approximately 10.5 years were well-suited for RNA-Seq compared to the intact tissues preserved for 1.5 years. Despite consistencies in RNA quantity, RNA integrity, amount of sequencing reads, and CRC gene signature, gene enrichment analysis revealed the decreased ribosome biogenesis, spliceosome and antifolate resistance pathways in the 2011–2012 group. Moreover, the FFPE tissues also showed no alteration in RNA quantity between the two periods, and the nCounter profiling demonstrated comparable CRC-specific gene counts in spite of the significant reduction of raw counts in the 2011–2012 group. Conclusions We report that FF tissues from CRC patients, stored for 10 years, are viable for whole transcriptome RNA-Seq, despite altered pathways such as ribosome biogenesis, spliceosome, and antifolate resistance. Moreover, 10-year-stored FFPE CRC tissues remain suitable for specific RNA profiling using the nCounter pan-cancer panel, despite a significant reduction in raw counts. These findings underscore the enduring contribution of biobanks to molecular research, highlighting their value a decade post-collection.

Objectives To explore and identify an optimal serum ferritin (SF) threshold level in diagnosing hemophagocytic lymphohistiocytosis (HLH) in Chinese children. Methods We conducted a retrospective study of 74 children with HLH admitted to Guangdong Women and Children Hospital between January 2015 and May 2021. Children in-hospital not diagnosed with HLH between January 2021 and May 2021 with a measurement of SF were enrolled as the non-HLH group. Patient charts were reviewed for SF levels upon admission and during hospitalization. A receiver operating characteristic (ROC) curve was utilized to determine the optimal cutoff value of SF for diagnosing childhood HLH. Results This study included a total of 74 children with HLH and 302 children with non-HLH diseases. The difference in SF values between the HLH and non-HLH groups was statistically significant (8,975 μg/L vs. 165.5 μg/L, p<0.001). An optimal SF cutoff value of 1,830 μg/L provided a sensitivity of 88 % and specificity of 79 % in confirming childhood HLH. The area under the curve (AUC) is 0.91 (95 % confidence interval 0.88–0.94, p<0.0001). Conclusions A serum ferritin level elevated above 1,830 μg/L might improve the specificity for HLH diagnosis in Chinese children.

Objectives To explore the usefulness of neutrophil-to-lymphocyte count ratio (NLR), procalcitonin (PCT), and interleukin-6 (IL-6) for the severity assessment of bacterial sepsis. Methods This study enrolled 100 patients with bacterial sepsis (disease group) who presented to Jinhua Central Hospital between March 2022 and March 2023 and 90 healthy individuals (control group). The patients were categorized into sepsis (64 cases), severe sepsis (18 cases), and septic shock (18 cases) groups according to the disease severity. The groups were compared in terms of the NLR, PCT, and IL-6, as well as the usefulness of these parameters, both alone and in combination, for the severity assessment of bacterial sepsis. Results The NLR, PCT, and IL-6 levels were significantly different among the three groups, with increasing values corresponding with disease aggravation. The area under the curve (AUC) values of the combinations of NLR, PCT, and IL-6 levels were higher than those of single markers. The sensitivity and AUC value of the combination of PCT and IL-6 levels were the highest (0.87), with a similar AUC value of the combination of NLR, PCT, and IL-6 (0.865); however, the specificity was significantly improved with the latter (0.938 vs. 0.859). Conclusions NLR, PCT, and IL-6 levels are significantly increased in bacterial sepsis, and the combination of PCT, and IL-6 levels can improve the sensitivity of the evaluation ability for severe sepsis, and is more economical.