High-Throughput and Automated Detection of HLA-B*27 Using the LabTurboTM AIO System.

The adoption of an automated system can decrease the hands-on time requirements in a clinical laboratory setting. For the detection of HLA-B*27, implementing a high-throughput and fully automated system has several advantages over using manual methods. Therefore, this study aimed to evaluate automation efficiency for the detection of HLA-B*27. Peripheral blood samples were obtained from 50 participants, and DNA was isolated from these samples. A Pharmigene PG27 detection kit was used for the qualitative detection of HLA-B*27. The performances of the semi-automated and fully automated LabTurboTM AIO systems in the detection of HLA-B*27 were compared. The mean absorbance (optical density) values for the MaelstromTM 8 and LabTurboTM AIO systems were found to be 1.88 and 1.9, respectively. The housekeeping gene was amplified and quantified using a real-time PCR assay across all DNA extracts to check the quality of the extracted human DNA. The results were expressed as the cycle threshold (Ct) values for all DNA extracts from both platforms. The mean Ct values for the Roche Cobas z480 and LabTurboTM AIO systems were found to be 22.7 and 20.4, respectively. This study demonstrated that the semi-automated method and the LabTurboTM AIO system yield consistent results for the detection of HLA-B*27. However, compared to the semi-automated method, the LabTurboTM AIO system provides standardized procedures, avoids manual handling, and improves turnaround time.

Transcriptome Profiling of Eutopic and Ectopic Endometrial Stromal Cells in Women with Endometriosis Based on High-Throughput Sequencing.

Endometriosis is a common gynecological disease that affects approximately 5-10% of reproductive-aged women. However, the etiology and pathophysiology of endometriosis are currently unclear. The objective of this study was to identify a potential pathogenic gene of endometriosis using RNA sequencing (RNA-seq) analysis. Human endometrial stromal cells were isolated from four patients receiving surgical treatment for endometriosis during laparoscopic surgery, and RNA-seq was used to examine differentially expressed genes (DEGs) in eutopic and ectopic endometrial stromal cells. The functional significance of the differentially expressed genes was analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A total of 1309 upregulated and 663 downregulated genes were identified through the analysis of the transcriptomes of eutopic and ectopic endometrial stromal cells. Furthermore, KEGG analysis indicated that these DEGs were mainly enriched in the PI3K-Akt signaling pathway, cytokine-cytokine receptor interaction, and MAPK signaling pathway. Our study identified differential gene expression in eutopic as compared to ectopic endometrial tissue stromal cells. We strongly believe that our findings can bring new insights into the underlying mechanisms of endometriosis. However, future research is necessary to clarify the roles of the identified genes.

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.