Assessment of the Clinical Utility of the FilmArrayTM Gastrointestinal Panel for Detecting Gastrointestinal Pathogens.

BACKGROUND: Gastrointestinal infections present a significant public health concern as they lead to diverse clinical presentations and healthcare challenges. The rapid and accurate identification of causative pathogens is imperative for effective patient management. This study aimed to assess the clinical utility of the FilmArrayTM Gastrointestinal (GI) Panel for detecting gastrointestinal pathogens. METHODS: Between November 1, 2022, and December 31, 2023, we analysed gastrointestinal specimens collected from a cohort of patients aged 21 to 91 at Asia University Hospital. These specimens were analyzed using the FilmArrayTM GI Panel. RESULTS: The study included 76 patients for whom the FilmArrayTM GI assay was conducted, with 40 (52.6%) showing positive results. Among the positive specimens, 23 (57.5%) had a single pathogen, while the remaining 17 (42.5%) had multiple pathogens. The remaining 36 (47.4%) specimens showed no pathogens. The overall positivity rate of the specimens was 52.6%. The most frequently detected pathogens included Salmonella, Clostridium difficile (toxin A/B), and Enteropathogenic Escherichia coli (EPEC). CONCLUSIONS: This study underscores the clinical value of the FilmArrayTM GI assay as a rapid and reliable tool for diagnosing gastrointestinal infections. Its capacity to detect multiple pathogens simultaneously enhances diagnostic accuracy and gives information to use in clinical decision-making. We strongly recommend its integration into clinical practice to expedite the diagnosis and management of gastrointestinal infections, ultimately leading to improved patient care and healthcare efficiency.

Performance Evaluation of the VIDAS® D-Dimer Exclusion™ II Assay and the Beckman Coulter D-Dimer Assay.

BACKGROUND: The breakdown of fibrinogen and fibrin during the process of fibrinolysis creates D-dimer. A D-dimer analysis is crucial for the diagnosis of pulmonary embolism, deep vein thrombosis, and disseminated intravascular coagulation. The aim of this study is to evaluate the validity of two different D-dimer assays. METHODS: To analyze the D-dimer, venous blood taken in a vacuette containing 3.2% sodium citrate was used. Peripheral whole blood specimens were collected from 89 subjects, and plasma was separated from these specimens. VIDAS® D-dimer Exclusion™ II and the Beckman Coulter D-dimer assays were used for the quantitative detection of D-dimer levels. The analytical performance of the two different D-dimer assays was compared. RESULTS: The plasma values of D-dimer ranged from 89.2 to 7,452.9 ng/mL (FEU) when tested on the VIDAS® platform and from 20 to 7,770 ng/mL (FEU) when tested on the Beckman Coulter platform. Our results showed that the agreement between the two methods was acceptable and Pearson's correlation coefficient (r) was 0.93 (p < 0.001). CONCLUSIONS: A high correlation exists between quantitative D-dimer measurements conducted with the VIDAS® D-dimer Exclusion™ II and the Beckman Coulter D-dimer assays.

Interference of Blood Glucose Testing by an Increase in Serum IgM Levels.

BACKGROUND: An absurd result of plasma glucose test caused by increased serum IgM levels. METHODS: Serum immunoglobulin levels are determined by using an immunoturbidimetric assay. Additionally, serial dilutions were performed to assess the absurd results. RESULTS: Our results showed that an increase in serum IgM levels induces errors in the measurement of the plasma glucose level. CONCLUSIONS: This study simply presents a message that both medical technologists and physicians need to be aware of this because improper results of blood glucose levels may result in aggressive and invasive patient management. Additionally, physicians may be led to wrong interpretation due to false levels of glucose. In fact, we do not know how often this situation accidentally occurs in the laboratory. Therefore, all medical technologists must stay alert to absurd and unusual test results and reconfirm the reason for an absurd result.

Optimizing Blood Culture Volumes by Implementing PDCA Cycle Management.

BACKGROUND: The aim of this study was to optimize the mean volume of blood drawn by nurses to a level that is recommended by our hospital through the implementation of PDCA cycle management. The purpose of the current study was to match the mean volumes of blood drawn with the volume recommended by the manufacturer. METHODS: The adequacy of blood volume in a bottle of aerobic blood culture per venipuncture was evaluated for every month from January 2021 to March 2022 by using the Becton Dickinson BD blood volume monitoring system. Furthermore, the study compared changes in the mean blood volumes before and after the PDCA cycle management was implemented. RESULTS: The mean blood volumes calculated for Q1 2021 (January 2021 to March 2021) before the PDCA cycle management was 6.3 mL per culture bottle. After PDCA cycle management was implemented, the mean blood volumes for Q1 2022 (January 2022 to March 2022) were calculated as 8.6 mL (p < 0.01). In addition, the positive culture rate increased from 13% to 15%. CONCLUSIONS: Implementing the PDCA cycle management can improve the mean blood culture volumes effectively and match the volume recommended by the manufacturer. Additionally, our study indicated that a higher blood volume yielded a culture rate that was significantly positive.

Evaluation of Friedewald's Formula for Plasma LDL-Cholesterol Estimation.

BACKGROUND: Low-density lipoprotein cholesterol (LDL-C) can contribute to atherosclerosis if it is oxidized within the walls of arteries. Therefore, LDL-C plays an important role in cardiovascular disease risk assessment and prevention. The current study aims to evaluate the validity of Friedewald's formula in the Taiwanese population. METHODS: In this analytical cross-sectional study, a data set containing 31,729 results was used and lipid profiles of all samples were measured using the Beckman Coulter AU680 clinical chemistry analyzer. This study was conducted from September 2016 to August 2019. RESULTS: The agreement between the direct and calculated LDL-C was significant with Pearson's correlation coefficient (r) of 0.904 (p < 0.001). Mean LDL-C levels were 99.3 ± 32.8 mg/dL and 95.3 ± 37.6 mg/dL for direct and calculated LDC-C, respectively. CONCLUSIONS: Good agreement was observed between direct and calculated LDC-C. Therefore, it can be concluded that Friedewald's formula is applicable in LDL-C estimation when the direct method is not affordable.

Retrospective Study of Lower Respiratory Tract Infections in the Intensive Care Unit Detected by the FilmArray Pneumonia Panel.

BACKGROUND: Lower respiratory tract infections (LRIs) are an important public health concern and a leading cause of death from infection worldwide. The current study aims to evaluate the distribution of viral and bacterial pathogens in lower respiratory tract specimens. METHODS: Between April 2022 and December 2022, specimens from lower respiratory tract from patients aged between 37 and 85 years in an intensive care unit (ICU) of Asia University Hospital were analysed by the FilmArrayTM pneumonia panel (PP) assay. RESULTS: There were 54 patients for whom the FilmArrayTM PP assay was analysed, and 25 (46.3%) of them showed positive results. Among the 54 specimens, 12 (22.2%, 12/54) had a single pathogen, 13 (24.1%, 13/54) had multiple pathogens, and 29 (53.7%, 29/54) had no pathogens. The overall positive rate of the specimens was 46.3% (25/54). CONCLUSIONS: The FilmArrayTM PP assay may act as a feasible diagnostic tool for LRIs in ICUs.

Detection of the SARS-CoV-2 D614G Mutation Using VirSNiP SARS-CoV-2 Spike D614G Assay.

BACKGROUND: At the end of 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly spread worldwide. Until recently, over 190 million cases and nearly four million deaths have been reported globally due to the virus. A point mutation in the SARS-CoV-2 spike protein, the D614G variant, was identified in late February 2020. Studies showed that the D614G mutation is related to higher infectivity. The current study was conducted to determine the validity of VirSNiP SARS-CoV-2 Spike D614G assay to detect the SARS-CoV-2 D614G mutation. METHODS: In this study, the detection of the SARS-CoV-2 D614G mutation was performed using VirSNiP SARS-CoV-2 Spike D614G assay (TIB Molbiol) and by Sanger sequencing. Two plasmids carrying A allele (wild-type) and G allele (mutant type) were designed using GenScript's OptimumGene gene design tool (GenScript Inc., Piscataway, NJ, USA). RESULTS: It was found that VirSNiP SARS-CoV-2 Spike D614G assay could detect the SARS-CoV-2 D614G mutation. The specific mutation is easily distinguishable in the melting peaks. All of the samples were confirmed using Sanger sequencing. CONCLUSIONS: In summary, it was shown that VirSNiP SARS-CoV-2 Spike D614G assay is a rapid, accurate, and reliable method for identifying the SARS-CoV-2 D614G mutation. In contrast, Sanger sequencing is time consuming and labor intensive. We strongly believe that VirSNiP SARS-CoV-2 Spike D614G assay will provide more information on the prevalence of the D614G mutation in Taiwanese populations.

Comparison of Manual and Automated Nucleic Acid Extraction Methods in Virus Transport Medium.

BACKGROUND: The implementation of an automated nucleic acid extraction system has many advantages over the manual methods. The purpose of this study was to evaluate the validity of two different methods for nucleic acid extraction in virus transport medium. METHODS: We collected 20 nasopharyngeal swabs in viral transport medium from the emergency department of the Asia University Hospital for the detection of SARS-CoV-2. The performance of the MaelstromTM 8 (Taiwan Advanced Nanotech) and the QIAamp Viral RNA Mini Kit (Qiagen) were compared for the extraction of nucleic acid from viral transport medium. The extracts were used for the validation of the RNA extraction procedures. The RNase P target was amplified in a one-step reverse transcription-quantitative PCR (RT-qPCR) reaction, as internal control for the extraction method. RESULTS: In this study, the agreement between the two methods was good and Pearson's correlation coefficient (r) was 0.919 (p < 0.001). The mean cycle threshold value of the two methods was 29.1. CONCLUSIONS: Overall, the performance values of the MaelstromTM 8 and the QIAamp Viral RNA Mini Kit were comparable to each other. In summary, the MaelstromTM 8 provides a standardized procedure, avoidance of sample-to-sample cross contaminations, is easy to use, improves turnaround time and requires less hands-on time as compared to the manual extraction method. The MaelstromTM 8 is more suitable for clinical laboratories that carry small or medium-sized samples for nucleic acid extraction.

Rapid Cobas Liat SARS-CoV-2 Assay in Comparison with the Laboratory-Developed Real-Time RT-PCR Test.

BACKGROUND: The objective of this study was to compare the validity of two different assays for the detection of SARS-CoV-2. METHODS: We collected 50 nasopharyngeal swabs in universal transport medium from the emergency department of Asia University Hospital for the detection of SARS-CoV-2 using reverse transcription-polymerase chain reaction (RT-PCR). The samples for the Liat SARS-CoV-2 influenza A/B test were stored at -70℃ after SARS-CoV-2 testing using the RT-PCR in order to assess method comparison. RESULTS: In this study, the Limit of detection (LOD) of the cobas Liat SARS-CoV-2 and influenza A/B nucleic acid test is 12 copies/µL and the assay obtained 100% positive agreement and negative percent agreement with RT-PCR. CONCLUSIONS: In summary, a prefect agreement exists between the detection of SARS-CoV-2 conducted with the cobas Liat SARS-CoV-2 and influenza A/B nucleic acid test and the RT-PCR. The cobas Liat SARS-CoV-2 and influenza A/B nucleic acid test is a reliable method for the detection of SARS-CoV-2, and it only requires 20 minutes to obtain the results. On the other hand, the cobas Liat SARS-CoV-2 and influenza A/B nucleic acid test is accurate, easy to use, and provides a faster turnaround time than testing performed in the high-throughput platform.

Analytical Performance of the VIDAS® BRAHMS Procalcitonin Assay and Beckman Coulter Unicel® DXI Procalcitonin Assay in a Stat Laboratory.

BACKGROUND: The objective of this study was to compare the validity of two different procalcitonin assays. METHODS: We collected 63 plasma samples from a stat laboratory. The plasma values of procalcitonin ranged from 0.01 to 98.1 µg/L when tested on the Access® platform and from < 0.05 to 98.5 µg/L when tested on the VIDAS® platform. The patients included 28 females ranging in age from 8 to 98 years of age (68 ± 22.6 years) and 35 males ranging in age from 35 to 90 years of age (69.2 ± 13.4 years). RESULTS: In this study, the agreement between the two methods was good and Pearson's correlation coefficient (r) was 0.989 (p < 0.001). CONCLUSIONS: In summary, a high correlation exists between quantitative procalcitonin measurements conducted with the VIDAS® BRAHMS and the Beckman Coulter Unicel® DXI assays. The VIDAS® BRAHMS procalcitonin assay is a reliable method for determining the levels of procalcitonin in plasma, but requires manual operation, hands-on technical expertise, and time. On the other hand, the Beckman Coulter Unicel® DXI assay is fully automated and may increase laboratory efficiency, and reduce the overall turnaround time.

A Comparison of Two Molecular Methods for Detecting CALR Mutations in Myeloproliferative Neoplasms.

BACKGROUND: Myeloproliferative neoplasms (MPN) are hematopoietic disorders characterized by abnormal proliferation of the myeloid lineage. Three classic subtypes are polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). These disorders are well known for their association with the JAK2 V617F mutation, in addition to mutations in MPL exon 10, and JAK2 exon 12. CALR mutations were detected in approximately 20% to 25% of patients with ET and PMF and not in patients with PV. Most CALR mutations were deletions and insertions in exon 9, which caused frameshift mutations. METHODS: This study included 60 Taiwanese patients with MPN. We identified CALR mutations in patients with MPN using the high-resolution melting (HRM) analysis. Additionally, the HRM analysis was compared with ipsogen CALR RGQ PCR. To confirm the results of HRM and ipsogen CALR RGQ PCR, sequencing analysis was also conducted all the samples. RESULTS: Up to 6.25% of CALR mutations were successfully detected in patients with MPN using HRM analysis. Eight out of 65 patients (12.3%) were positive for the presence of CALR mutation, including p.L367fs*46 and p.K385fs*47. The results proved 100% comparable to those obtained using ipsogen CALR RGQ PCR. CONCLUSIONS: The HRM analysis and ipsogen CALR RGQ PCR are feasible and reliable techniques for the detection of CALR mutation. Furthermore, HRM offers several benefits, for example, it is time-saving, inexpensive, and does not require many personnel.

Gender differences in geriatric syndromes as mental illness and nervous system diseases in hospitalized Thai older patients.

BACKGROUND: Older persons are affected by mental and neurological disorders differently, and gender plays a significant role influencing geriatric disorder differentiation. Accordingly this study characterized gender differences in geriatric syndromes among hospitalized elderly Thai patients. METHODS: Probabilities of disease occurrence reflecting gender differences were calculated using historical data obtained from the Ministry of Public Health website, Thailand. We selected older patients aged 60 years and above admitted to inpatient departments in public hospitals with mental disorders and nervous system diseases from 2014 to 2018, counting over 160 000 cases each year. Descriptive statistics and odds ratios (ORs) were used to analyse and demonstrate gender differences. RESULTS: Compared to older females, older males had higher occurrences of four mental disorders revealed by OR and 95% confidence interval (CI) values: substance abuse (5.74, 5.08-6.49), alcohol use (5.66, 5.44-5.89), behavioural problems (1.34, 1.31-1.37), and schizophrenia (1.10, 1.06-1.14). Lower incidences for older males were seen in the same values for three mental disorders: neurotic issues (0.46, 0.44-0.49), mood disorders (0.58, 0.56-0.60), and dementia (0.91, 0.88-0.94). For neurological disorders, men had similar higher incidences for epilepsy (1.67, 1.63-1.72), cerebral palsy (1.61, 1.57-1.65), nervous system inflammatory diseases (1.53, 1.46-1.60), ischaemic attacks (1.42, 1.36-1.48), miscellaneous other nervous disorders (1.20; 1.18-1.22), and Parkinson's disease (1.15, 1.12-1.19). By contrast, older men had lower incidences of multiple sclerosis (0.55, 0.35-0.86), migraines (0.66, 0.62-0.70), and Alzheimer's disease (0.75, 0.71-0.78). CONCLUSION: Accurate characterization of gender differences in geriatric syndromes can better inform policies for appropriate early detection and prevention, and contribute to the development of treatment and intervention for various issues affecting elderly men and women's health.

Identification of a Case with Heterozygous Mutations in the FUT1 Gene Leading to a Para-Bombay Phenotype.

BACKGROUND: A case of a para-Bombay phenotype caused by a compound heterozygous mutation in the FUT1 gene was identified in this study. METHODS: We performed an agglutination examination of anti-H serum and secretor status to assess the presence of soluble blood group substances. Genotyping of ABO and FUT1 genes was also performed. RESULTS: Our results showed the presence of A and H antigens in saliva. Based on these results, the patient in the present case was diagnosed with the para-Bombay A phenotype. Direct DNA sequencing of the ABO gene indicated A1v/O1vgenotype. FUT1 gene sequence analysis revealed that the patient harbored the compound heterozygous mutation, c.881_882delTT (p.Phe294Cysfs*40) and c.658C>T (p.Arg220Cys). CONCLUSIONS: Improper identification of this phenotype may cause inappropriate transfusions because this particular blood group may be mislabeled as group O. Therefore, blood bank staff should be well trained to solve the discrepancy between cell and serum grouping in the para-Bombay phenotype.

Analytical Performance of the VIDAS® High-Sensitivity Troponin I Assay and the Beckman Coulter Unicel® DXI AccuTnI+3 Assay in a Stat Laboratory.

BACKGROUND: The aim of this study was to compare the validity of two different cTnI assay methodologies. METHODS: We collected 82 plasma samples from a stat laboratory. The plasma values of cTnI ranged from 0.012 to 29.715 ng/mL when tested on the Access® platform and from 4.5 to >40,000 ng/L when tested on the VIDAS platform. The patients included 34 females ranging in age from 49 to 100 years of age [76.7 ± 12 years] and 48 males ranging from 29 to 97 years of age [69.7 ± 12 years]. RESULTS: Our results showed that the correlation between the two troponin results was r2 = 0.9836 (p < 0.001). In this study, the kappa statistic (0.89) indicated a high degree of agreement between the VIDAS® High-sensitivity Troponin I assay and the Beckman Coulter Unicel® DXI AccuTnI+3 assay. CONCLUSIONS: In summary, the VIDAS® High-sensitivity Troponin I assay is a reliable and feasible method for determining the levels of cTnI in plasma, but it requires manual operation, hands-on technical expertise, and time.

Heterozygous FUT1 Mutations Causing a Para-Bombay Phenotype.

BACKGROUND: To study a case of para-Bombay phenotype caused by compound heterozygous mutation of the FUT1 gene. METHODS: We performed an agglutination examination to anti-H serum. Secretor status was determined in order to assess the presence of soluble blood group substances. Genotyping of ABO and FUT1 genes were also performed. RESULTS: Our results showed the presence of A and H antigens in the saliva. Based on these results, the patient in the present case was diagnosed to have a para-Bombay A phenotype. Direct DNA sequencing of the patient's ABO gene indicated A1v/O1vgenotype. FUT1 gene sequence analysis revealed that the patient harbored the compound heterozygous mutations c.881_882delTT (p.Phe294Cysfs) and c.551_552delAG (p.Glu184Valfs). CONCLUSIONS: In summary, our findings support that the occurrence of a heterozygous mutation in FUT1, 547delAG/880delTT, is the most common mutation in Taiwanese.

Molecular characterization of oral squamous cell carcinoma using targeted next-generation sequencing.

OBJECTIVES: Many genetic factors play an important role in the development of oral squamous cell carcinoma. The aim of this study was to assess the mutational profile in oral squamous cell carcinoma using formalin-fixed, paraffin-embedded tumors from a Taiwanese population by performing targeted sequencing of 26 cancer-associated genes that are frequently mutated in solid tumors. METHODS: Next-generation sequencing was performed in 50 formalin-fixed, paraffin-embedded tumor specimens obtained from patients with oral squamous cell carcinoma. Genetic alterations in the 26 cancer-associated genes were detected using a deep sequencing (>1000X) approach. RESULTS: TP53, PIK3CA, MET, APC, CDH1, and FBXW7 were most frequently mutated genes. Most remarkably, TP53 mutations and PIK3CA mutations, which accounted for 68% and 18% of tumors, respectively, were more prevalent in a Taiwanese population. Other genes including MET (4%), APC (4%), CDH1 (2%), and FBXW7 (2%) were identified in our population. CONCLUSIONS: In summary, our study shows the feasibility of performing targeted sequencing using formalin-fixed, paraffin-embedded samples. Additionally, this study also reports the mutational landscape of oral squamous cell carcinoma in the Taiwanese population. We believe that this study will shed new light on fundamental aspects in understanding the molecular pathogenesis of oral squamous cell carcinoma and may aid in the development of new targeted therapies.

Comprehensive analysis of UGT1A1 polymorphisms through high-resolution melting analysis and DNA sequencing.

BACKGROUND: Uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) is a key conjugating enzyme of bilirubin and the anti-tumor medication irinotecan. Comprehensive analysis of UGT1A1 gene polymorphisms may provide benefit by predicting pharmacokinetics and outcomes of treatment with irinotecan and certain antiviral medications. METHODS: A high-resolution melting (HRM) analysis was designed to characterize the UGT1A1 gene. Genomic DNA from 110 healthy subjects was extracted from peripheral blood samples. The promoter and 11 exons from UGT1A1 were screened by HRM, and all results were subsequently confirmed by direct DNA sequencing. RESULTS: HRM analysis readily identified UGT1A1 gene mutations. We identified 5 different known variants of UGT1A1 including c.211 G > A; G71R, c.686 C > A; P229Q, c.1091 C > T; c.-3279 T > G; and c.-3156 G > A in 110 normal Taiwanese individuals. We also identified 8 new sequence variants, namely, c.-3296 C > T; c.43 C > A; c.45 G > A; c.234 G > A; c.577 G > A; c.614 C > T; c.1011 T > C; and c.1352 C > T. Each UGT1A1 variant was easily identifiable by differences in curves plotted from HRM data. CONCLUSIONS: HRM analysis was rapid, accurate, and economical for screening UGT1A1 gene mutations.

Correlations of sST2 and Gal-3 with Cardiothoracic Ratio in Patients with Chronic Kidney Disease.

Chronic kidney disease (CKD) frequently correlates with cardiovascular complications. Soluble suppression of tumorigenicity 2 (sST2) and Galectin-3 (Gal-3) are emerging as cardiac markers with potential relevance in cardiovascular risk prediction. The cardiothoracic ratio (CTR), a metric easily obtainable from chest radiographs, has traditionally been used to assess cardiac size and the potential for cardiomegaly. Understanding the correlation between these cardiac markers and the cardiothoracic ratio (CTR) could provide valuable insights into the cardiovascular prognosis of CKD patients. This study aimed to explore the relationship between sST2, Gal-3, and the CTR in individuals with CKD. Plasma concentrations of sST2 and Gal-3 were assessed in a cohort of 123 CKD patients by enzyme-linked immunosorbent assay (ELISA). On a posterior-to-anterior chest X-ray view, the CTR was determined by comparing the widths of the heart to that of the thorax. The mean concentration of sST2 in the study participants ranged from 775.4 to 4475.6 pg/mL, and the mean concentration of Gal-3 ranged from 4.7 to 9796.0 ng/mL. Significant positive correlations were observed between sST2 and the CTR (r = 0.291, p < 0.001) and between Gal-3 and the CTR (r = 0.230, p < 0.01). Our findings indicate that elevated levels of sST2 and Gal-3 are associated with an increased CTR in CKD patients. This relationship may enable better cardiovascular risk evaluation for CKD patients. Further studies are warranted to explore the clinical implications of these associations.