1.Construction of Perimenopausal Depression Animal Models and Mechanism of Action of Traditional Chinese Medicine: A Review
Xiaoting LI ; Shouzhu XU ; Jun KE ; Zhan ZHANG ; Bao XIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):258-267
Perimenopausal depression (PMD) is an affective disorder that occurs in women during the transition from sexual maturity to old age. It can induce various complications, such as insomnia and cognitive decline. The etiology of PMD is complex. Although multiple hypotheses have been proposed, there is still no unified theory that fully explains its pathogenesis. Research into its mechanisms relies heavily on animal experiments, and establishing reliable animal models is crucial for experimental studies. Appropriate animal models can better simulate human pathophysiological states, rapidly evaluate the efficacy and safety of drugs and intervention methods, grasp the essence of the disease, and uncover its intrinsic connections, thereby exploring more advanced intervention strategies. However, there is a lack of systematic review and summarization of literature related to model construction. Additionally, traditional Chinese medicine (TCM), adhering to the principles of ''syndrome differentiation and treatment'' and ''holistic concept'', has shown significant efficacy in treating PMD. In recent years, research exploring and analyzing its therapeutic mechanisms has been increasing. Therefore, to gain a clearer and more comprehensive understanding of PMD animal modeling methods and the mechanisms of TCM, this paper reviewed Chinese and English literature on PMD animal models and mechanisms of TCM in PMD treatment. It summarized the construction methods of single-factor and multi-factor PMD models, and discussed the advantages and disadvantages of each modeling approach. Furthermore, it delved into the mechanisms of TCM intervention in PMD, revealing that TCM formulas primarily exert their effects by regulating the hypothalamic-pituitary-adrenal axis, hypothalamic-pituitary-ovarian axis, gut-brain axis, cell signaling pathways, neural circuits, hormone levels, and neurotransmitter levels. This review aims to provide a reference for future research in this field. In summary, by summarizing the progress in the methods for PMD animal model construction and the mechanisms of TCM, the paper seeks to offer new insights into the mechanistic research of TCM intervention in PMD.
2.Standards for the Application of Hemodynamic Monitoring Technology in Critical Care
Hua ZHAO ; Hongmin ZHANG ; Xin DING ; Huan CHEN ; Jun DUAN ; Wei DU ; Bo TANG ; Yuankai ZHOU ; Dongkai LI ; Xinchen WANG ; Cui WANG ; Gaosheng ZHOU ; Xiaoting WANG
Medical Journal of Peking Union Medical College Hospital 2026;17(1):73-85
With the rapid advancement of hemodynamic indices and monitoring technologies, their classification methods and application processes have become increasingly complex. Currently, no unified standard hasbeen established, making it difficult to fully meet the clinical requirements for hemodynamic management. To assist in hemodynamic monitoring assessment and therapeutic decision-making in critically ill patients, the Critical Hemodynamic Therapy Collaborative Group, in conjunction with the Critical Ultrasound Study Group, has jointly developed the Standard for the Application of Hemodynamic Monitoring Techniques in Critical Care. The first part of this standard systematically categorizes hemodynamic indicators into flow indicators, pressure and its derivative indicators, and tissue perfusion indicators, while elaborating on the clinical application of each. The second part establishes a standardized clinical implementation pathway for hemodynamic monitoring. It proposes a tiered monitoring strategy-comprising basic, advanced, indication-specific, and special scenario monitoring-tailored to different clinical settings. It emphasizes the central role of critical care ultrasound across all levels of monitoring and establishes hemodynamic assessment standards for organs such as the brain, kidneys, and gastrointestinal tract. This standard aims to provide a unified framework for clinical practice, teaching, training, and research in critical care medicine, thereby promoting standardized development within the discipline.
3.Evaluation of the anticoagulant effect of nafamostat mesylate in continuous veno-venous hemofiltration with different dilution methods for uremic patients
Li SHEN ; Yao ZHANG ; Jun WANG ; Hong ZHU ; Yong QIN ; Yuewu TANG ; Ni DU
China Pharmacy 2026;37(3):350-355
OBJECTIVE To evaluate the anticoagulant efficacy and safety of nafamostat mesylate (NM) in the treatment of uremic patients at high risk of bleeding undergoing continuous veno-venous hemofiltration (CVVH) with different methods (pre- dilution and post-dilution). METHODS A total of 130 uremic patients at high risk of bleeding who underwent CVVH treatment in the nephrology department of Chongqing University Three Gorges Hospital from July 2023 to September 2024 were selected. They were divided into pre-dilution group and post-dilution group according to the random number table method, with 65 cases in each group. Both groups of patients received CVVH treatment under NM anticoagulation. The pre-dilution group adopted the pre-dilution replacement method, while the post-dilution group adopted the post-dilution replacement method. The coagulation, pressure, and usage duration of the filter and dialysis circuit venous reservoirs were compared between the two groups. The changes in prothrombin time (PT), prothrombin time-international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), and fibrinogen (FIB) in the peripheral venous blood before the heparin pump and after the filter at 1, 4 and 7 h of CVVH treatment, as well as 20 min after the end of treatment, were compared between the two groups. The single-compartment urea clearance rate (spKt/V), β2-microglobulin (β2-MG) clearance rate and the incidence of adverse reactions were duni2007@foxmail.com compared between the two groups. RESULTS Both the pre-dilution and post-dilution groups had 60 patients who completed the study. The incidence of grade Ⅱ-Ⅲ coagulation of the filter and venous reservoirs, as well as the number of patients with transmembrane and venous pressure alarm intervention in the post- dilution group were significantly higher or more than those in the pre-dilution group (P<0.05), while usage time of the filter and the pipeline in the post-dilution group was significantly shorter than that in the pre-dilution group (P<0.05). The APTT values before the heparin pump as well as PT and APTT values after the filter at 1 h, 4 h, and 7 h of CVVH treatment in the post-dilution group were significantly higher than those in the pre-dilution group (P<0.001). There were no significant differences in PT, PT- INR, APTT and FIB between the two groups of patients 20 min after the end of treatment (P>0.05). The spKt/v and β2-MG clearance rates in the post-dilution group were significantly higher than those in the pre-dilution group (P<0.001). There was no significant difference in the incidence of adverse reactions between the two groups (P>0.05). CONCLUSIONS When NM is used as an anticoagulant in the CVVH treatment of uremic patients at high risk of bleeding, compared with the pre-dilution treatment method, the post-dilution treatment method has a higher incidence of filter and dialysis tubing venous reservoir, a shorter usage time of the filter and pipeline, and a greater impact on extracorporeal coagulation, but has a higher solute clearance rate. Clinically, different dilution methods can be selected according to the different treatment needs of patients.
4.Body image disturbance status in AS patients and analysis of its influencing factors
Min NIU ; Jingman YUAN ; Liya MA ; Hao XU ; Jun LI ; Meixi YAN ; Xinru DU ; Hanhui MA ; Xichao YANG
Journal of Public Health and Preventive Medicine 2026;37(1):158-162
Objective To understand the status of body image disturbance and its influencing factors in patients with ankylosing spondylitis (AS), so as to provide a scientific basis for the clinical management of AS. Methods A total of 353 AS patients admitted from January 2022 to December 2024 were selected as research subjects. Chinese version of Body Image Disturbance Questionnaire (BIDQ) was used to investigate the body image disturbance in AS patients. Single factor analysis was performed by t test and analysis of variance, and multiple factors were analyzed by multivariate linear regression. Results The total score of BIDQ in 342 AS patients was (25.01±4.22). Multivariate linear regression analysis results showed that self-paid medical expense, nighttime VAS score and negative emotion PANAS score could positively predict body image disturbance in AS patients (standardized regression coefficient=0.413, 0.413, 0.460, P<0.05), and PSSS score, positive emotion PANAS score and exercise management CDSSM score could negatively predict body image disturbance (standardized regression coefficient=-0.245, -0.134, -0.247, P<0.05). Conclusion The body image disturbance in AS patients is worthy of clinical attention. Nighttime pain, negative emotion and self-paid medical treatment can increase the risk of body image disturbance. Positive emotion, social support and high self-management level of exercise behavior can reduce the formation of body image disturbance, which can provide new ideas for clinical management of AS patients.
5.Research on the screening efficiency of Thalassemia based on an automated evaluation software.
Jun HU ; Huan LIANG ; Limei DUAN ; Jianqiang GAO
Chinese Journal of Medical Genetics 2026;43(4):281-287
OBJECTIVE:
To explore the efficacy of a Thalassemia risk assessment software for the screening of thalassemia mutation carriers and distribution of thalassemia genotypes detected by screening.
METHODS:
A total of 6 040 individuals were evaluated at Leshan Maternal and Child Health Care Hospital between 2022 and 2024 using the commonly used clinical thalassemia risk assessment method and the thalassemia screening software, respectively, and the performance indicators of the two methods were compared and analyzed against the result of thalassemia gene testing. This study was approved by the Ethics Committee of our hospital (Ethics No.: LfyLL[2022]005).
RESULTS:
The high-risk rate by the thalassemia screening software was 11.19%, with a sensitivity of 95.12%, specificity of 93.28%, positive predictive value of 43.20%, negative predictive value of 99.72%, and the area under the ROC curve (AUC) was 0.942. The thalassemia gene detection rate of the high-risk samples screened was 4.83%. The high-risk screening rate of the conventional method was 2.50%, with a sensitivity of 51.22%, specificity of 93.28%, positive predictive value of 80.79%, negative predictive value of 97.40%, and the AUC was 0.754. The thalassemia gene detection rate of the high-risk samples was 2.02%.
CONCLUSION
The software can effectively detect thalassemia carriers and significantly reduce the missed detection compared with conventional method, thereby significantly improve the efficacy of screening.
Humans
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Thalassemia/diagnosis*
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Software
;
Female
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Genetic Testing/methods*
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Male
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Mutation
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Adult
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Genotype
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ROC Curve
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Risk Assessment
6.Xuefu Zhuyutang Ameliorates Metabolic-associated Fatty Liver Disease via AMPK Signaling Pathway
Ming HAN ; Ying ZHANG ; Lingya KONG ; Jun DAI ; Ting ZHANG ; Zhihong MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):1-12
ObjectiveTo investigate the therapeutic mechanism of Xuefu Zhuyutang (XFZYT) for metabolic-associated fatty liver disease (MAFLD) through integrated network pharmacology and animal experiments. MethodsNetwork pharmacology was utilized to predict the core components, key therapeutic targets, and signaling pathways of XFZYT in the treatment of MAFLD. For animal experiments, a rat model of MAFLD was established by feeding a high-cholesterol diet for 4 weeks. Intervention was then administered with low-dose (2 g·kg-1) and high-dose (4 g·kg-1) XFZYT for 2 weeks. Biochemical assays were performed to measure the serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL). In addition, the activities of superoxide dismutase (SOD) and catalase (CAT) and levels of malondialdehyde (MDA) and glutathione (GSH) in the serum were measured. The same way was adopted to measure the levels of TC and TG in the liver tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the serum levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor-alpha (TNF-α). Histopathological evaluations included hematoxylin and eosin (HE) staining for liver tissue morphology, Oil Red O staining for lipid deposition, and dihydroethidium (DHE) probe staining for reactive oxygen species (ROS) levels. Western blot analysis was conducted to assess the protein levels of AMP-activated protein kinase (AMPK), phosphorylated (p)-AMPK, nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), nuclear factor-kappa B (NF-κB), and p-NF-κB in the liver tissue. Untargeted metabolomics analysis of the serum was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). ResultsNetwork pharmacology analysis predicted 155 potential targets of XFZYT for MAFLD treatment, with core targets including signal transducer and activator of transcription 3 (STAT3), protein kinase B1 (Akt1), TNF, and IL-6. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment primarily implicated the AMPK signaling pathway. Animal experiments demonstrated that compared with the normal group, the model group exhibited dyslipidemia, hepatic function impairment, pronounced hepatic lipid deposition, and inflammatory manifestations, with elevated serum levels of AST, ALT, TC, TG, LDL, and MDA (P<0.05), reduced HDL and GSH levels plus decreased SOD and CAT activities (P<0.05), downregulated protein levels of Nrf2, HO-1, and p-AMPK (P<0.05), and upregulated protein level of p-NF-κB (P<0.05) in the liver tissue. Compared with the model group, XFZYT intervention groups showed significant amelioration of dyslipidemia and hepatic function impairment, markedly reduced hepatic lipid deposition and inflammatory cell infiltration, decreased serum levels of AST, ALT, TC, TG, LDL, and MDA (P<0.05), increased HDL and GSH levels plus enhanced SOD and CAT activities (P<0.05), upregulated protein levels of Nrf2, HO-1, and p-AMPK (P<0.05), and downregulated protein level of p-NF-κB (P<0.05). Serum metabolomics revealed 511 differentially expressed metabolites (231 upregulated and 280 downregulated) between normal and model groups, while XFZYT groups versus model group showed 94 differential metabolites (51 upregulated and 43 downregulated). Among them, 11 metabolites displayed the most significant alterations, with enriched pathways including glycerolipid metabolism, cholesterol metabolism, and insulin resistance, multiple of which demonstrated AMPK association. ConclusionXFZYT alleviates MAFLD by regulating the AMPK signaling pathway and associated metabolic networks.
7.Regulatory Mechanisms of miRNA in Hepatocellular Carcinoma and Current Status of Traditional Chinese Medicine Intervention: A Review
Shiyi LI ; Hao CHENG ; Chunyan JI ; Jun SUN ; Juan XUE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):323-332
Hepatocellular carcinoma(HCC), as one of the common malignant tumours, has seen a continuous rise in incidence and mortality worldwide, posing a serious threat to human health. However, traditional treatments have certain limitations, therefore, the exploration of new therapeutic strategies is particularly urgent. In recent years, with in-depth research on the regulatory mechanisms of microRNA(miRNA) in tumour occurrence and development, it has become new targets for HCC diagnosis and treatment. As a traditional treatment method, Chinese medicine, due to its multi-component, multi-pathway, and multi-target overall regulatory characteristics, shows broad prospects in treating HCC by regulating miRNAs. Accordingly, this paper reviews recent studies on the role of miRNAs in HCC and research advances in traditional Chinese medicine interventions, finding that various miRNAs play key roles in HCC cell cycle regulation, proliferation and apoptosis, invasion and metastasis, immune microenvironment, and drug resistance. It summarises how active ingredients, extracts, medicinal pairs, and formulas of Chinese medicine act on specific miRNAs to regulate their downstream target gene expression, affecting the malignant behaviour of HCC cells and exerting anti-cancer effects. This study aims to provide a theoretical basis for miRNAs as potential biomarkers and therapeutic targets for HCC, as well as to offer new ideas for developing miRNA-based targeted Chinese medicine therapies.
8.Effects of LINC02086 on proliferation, migration and invasion of gastric cancer cells by regulating Wnt/β-catenin pathway mediated M2 polarization of macrophages
Jun LI ; Yafei BU ; Jie CHEN ; Bo DING ; Lei WANG
Acta Universitatis Medicinalis Anhui 2026;61(2):192-201
ObjectiveTo investigate the effect and mechanism of long intergenic non-coding RNA02086 (LINC02086) overexpression mediated macrophage polarization on the proliferation, migration and invasion of gastric cancer cells. MethodsThe expression levels of LINC02086 in the human gastric epithelial cell line GES-1 and human gastric cancer cell lines HCG-27, NCI-N87, and AGS were determined by qRT-PCR. Human acute monocytic leukemia cells (THP-1) were induced to differentiate into M0 macrophages using phorbol 12-myristate 13-acetate (PMA). HGC-27 cells were infected with either LINC02086 overexpression lentivirus (OE-LINC02086) or its negative control lentivirus (Vector), and the culture supernatants were collected as conditioned medium (CM1). M0 macrophages were co-cultured with the infected HGC-27 cells, and the resulting supernatants were designated as conditioned medium 2 (CM2). M0 macrophages were treated with CM1 alone or in combination with Wnt/β-catenin pathway inhibitor IWR-1, forming the Vector+CM1, OE-LINC02086+CM1, and OE-LINC02086+CM1+IWR-1 groups, respectively. Flow cytometry was used to detect mannose receptor C-type 1 (CD206) expression, and qRT-PCR was employed to measure mRNA levels of interleukin-10 (IL⁃10), transforming growth factor-β (TGF⁃β), vascular endothelial growth factor (VEGF), and chemokine ligand 22 (CCL22). Western blot was performed to evaluate protein expression of CD206, VEGF, and key components of the Wnt/β-catenin pathway—Wnt family member 3a (Wnt3a), glycogen synthase kinase-3β (GSK-3β), and β-catenin. HGC-27 cells were treated with CM2 alone or combined with IWR-1, establishing the Vector+CM2, OE-LINC02086+CM2, and OE-LINC02086+CM2+IWR-1 groups. CCK-8 assay was used to evaluate cell proliferation, and Transwell assays were conducted to assess migration and invasion capabilities. ResultsCompared with GES-1 cells, the expression levels of LINC02086 were upregulated in HCG-27, NCI-N87, and AGS cells (P < 0.05), with the smallest increase observed in HCG-27 cells. Compared with Vector+CM1 group, the level of CD206 and the expression levels of IL⁃10, TGF⁃β, VEGF and CCL22 mRNA in macrophages stimulated by OE-LINC02086+CM1 increased (P<0.05). Meanwhile, the expression levels of Wnt3a and β-catenin proteins in cells increased (P<0.05), and the expression level of GSK-3β protein decreased (P<0.05). However, co-treatment with IWR-1 markedly reversed the promoting effects of LINC02086 overexpression on the expression of M2 polarization markers, including CD206, IL⁃10, and TGF⁃β mRNA, in macrophages (P<0.05), as well as its activation of the Wnt/β-catenin signaling pathway (P<0.05). Compared with Vector+CM2 group, HGC-27 cells infected with OE-LINC02086+CM2 had increased proliferation activity and increased number of migration and invasion cells (P<0.05). However, the combined intervention of IWR-1 significantly reversed the promotion of LINC02086 overexpression on the proliferation, migration and invasion of HGC-27 cells (P<0.05). ConclusionLINC02086 overexpression promotes the proliferation, migration and invasion of gastric cancer cells by activating Wnt/β-catenin pathway to mediate M2 polarization of macrophages.
9.Diagnostic value of serum Mac-2 binding protein for the severity of schistosomiasis-induced liver fibrosis
Jun WU ; Meiqun LUO ; Shuying XIE ; Ronghua ZHU ; Hui XU ; Long TANG ; Fei HU ; Sheng DING
Chinese Journal of Schistosomiasis Control 2026;38(1):38-43
Objective To evaluate the value of serum Mac-2 binding protein (M2BP) for assessment of the severity of schisto somiasis-induced liver fibrosis, so as to provide insights into non-invasive diagnosis and disease surveillance of liver fibrosis caused by schistosomiasis. Methods A total of 234 individuals with a history of Schistosoma japonicum infection were sampled from Xinhua Village, Lushan City, Jiangxi Province from 2019 to 2020, and 234 serum samples were collected from all participants. All participants received B-ultrasound examinations of the liver. Serum samples were categorized into four groups (grades 0, Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis groups) according to B-ultrasound examination results, and then, each group was randomly divided into a receiver operating characteristic (ROC) curve group and an efficacy assessment group at a ratio of 7∶3. Serum M2BP concentration was measured in four groups using the enzyme-linked immunosorbent assay (ELISA), and differences in serum M2BP concentrations were compared with analysis of variance and Spearman correlation analysis. Serum M2BP concentration was subjected to ROC curve analysis among individuals with different grades of schistosomiasis-induced liver fibrosis in the ROC curve group to determine the optimal diagnostic threshold of M2BP concentration at different fibrosis grades, and the area under the ROC curve (AUC) was calculated to evaluate the diagnostic performance. The diagnostic accuracy was verified by comparing the accordance rate and Kappa consistency test in the efficacy assessment group. Results Among 234 serum samples, there were 79 samples with grade 0 schistosomiasis-induced liver fibrosis, 87 samples with Grade Ⅰ, 46 samples with Grade Ⅱ and 22 samples with Grade Ⅲ according to the B-ultrasound examinations. The mean serum M2BP concentrations were (0.40 ± 0.31) [95% confidence interval (CI): (0.33, 0.47)], (0.64 ± 0.48) [95% CI: (0.53, 0.74)], (1.76 ± 0.58) [95% CI: (1.59, 1.93)] μg/mL and (2.56 ± 0.93) [95% CI: (2.14, 2.97)] μg/mL in the four groups, respectively (F = 150.796, P < 0.001), and the severity of schistosomiasis-induced liver fibrosis significantly positively correlated with serum M2BP concentration (rs = 0.715, P < 0. 001). The sample sizes of grades 0, Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis sera were randomly allocated as follows: 55 versus 24, 61 versus 26, 32 versus 14, and 15 versus 7 in the ROC curve and efficacy assessment groups, respectively, and the serum M2BP concentrations were (0.39 ± 0.29) μg/mL and (0.42 ± 0.36) μg/mL (F = 0.196, P > 0.05), (0.59 ± 0.47) μg/mL and (0.75 ± 0.51) μg/mL (F = 1.967, P > 0.05), (1.73 ± 0.59) μg/mL and (1.85 ± 0.57) μg/mL (F = 0.417, P > 0.05), and (2.46 ± 0.64) μg/mL and (2.76 ± 1.41) μg/mL (F = 0.491, P > 0.05), respectively. ROC curve analysis showed that the optimal diagnostic thresholds of serum M2BP concentration were 0.347 86 μg/mL (AUC = 0.635, P < 0.05), 1.188 83 μg/mL (AUC = 0.938, P < 0.000 1) and 2.021 21 μg/mL (AUC = 0.821, P < 0.000 1) for grade Ⅰ, Ⅱ and Ⅲ schistosomiasis-induced liver fibrosis. In addition, the accordance rates between the optimal diagnostic threshold of serum M2BP and B-ultrasound examinations for predicting grade Ⅰ, Ⅱ and Ⅲ schistosomiasis-induceed liver fibrosis were 69.23%, 85.71% and 71.43% (χ2 = 1.340, P > 0.05), and the overall Kappa consistency test showed moderate consistency [Kappa = 0.608, 95% CI: (0.428, 0.788); Z = 6.609, P < 0.000 1]. Conclusions Serum M2BP may serve as a potential biomarker for assessing moderate to advanced schistosomiasis-induced liver fibrosis; however, its diagnostic value for early-stage schistosomiasis-induced liver fibrosis remains limited.
10.Genomic variant surveillance of SARS-CoV-2 positive specimens using a direct PCR product sequencing surveillance (DPPSS) method.
Nicole Ann L. Tuberon ; Francisco M. Heralde III ; Catherine C. Reportoso ; Arturo L. Gaitano III ; Wilmar Jun O. Elopre ; Kim Claudette J. Fernandez
Acta Medica Philippina 2026;60(1):57-68
BACKGROUND AND OBJECTIVE
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as the causative agent of COVID-19 has significantly challenged the public health landscape in late 2019. After almost 3 years of the first ever SARS-CoV-2 case, the World Health Organization (WHO) declared the end of this global health emergency in May 2023. Although, despite the subsequent drop of COVID-19 cases, the SARS-CoV-2 infection still exhibited multiple waves of infection, primarily attributed to the appearance of new variants. Five of these variants have been classified as Variants of Concern (VOC): Alpha, Beta, Gamma, Delta, and the most recent, Omicron. Therefore, the development of methods for the timely and accurate detection of viral variants remains fundamental, ensuring an ongoing and effective response to the disease. This study aims to evaluate the feasibility of the application of an in-house approach in genomic surveillance for the detection of SARS-CoV-2 variants using in silico designed primers.
METHODSThe primers used for the study were particularly designed based on conserved regions of certain genes in the virus, targeting distinct mutations found in known variants of SARS-CoV-2. Viral RNA extracts from nasopharyngeal samples (n=14) were subjected to quantitative and qualitative tests (Nanodrop and AGE). Selected samples were then analyzed by RT-PCR and amplicons were submitted for sequencing. Sequence alignment analysis was carried out to identify the prevailing COVID-19 variant present in the sample population.
RESULTSThe study findings demonstrated that the in-house method was able to successfully amplify conserved sequences (spike, envelope, membrane, ORF1ab) and enabled identification of the circulating SARS-CoV-2 variant among the samples. Majority of the samples were identified as Omicron variant. Three out of four designed primers effectively bound into the conserved sequence of target genes present in the sample, revealing the specific SARSCoV-2 variant. The detected mutations characterized for Omicron found in the identified lineages included K417N, S477N, and P681H which were also identified as mutations of interest. Furthermore, identification of the B.1.448 lineage which was not classified in any known variant also provided the potential of the developed in-house method in detecting unknown variants of COVID-19.
CONCLUSIONAmong the five VOCs, Omicron is the most prevalent and dominant variant. The in-house direct PCR product sequencing surveillance (DPPSS) method provided an alternative platform for SAR-CoV-2 variant analysis which is accessible and affordable than the conventional diagnostic surveillance methods and the whole genome sequencing. Further evaluation and improvements on the oligonucleotide primers may offer significant contribution to the development of a specific and direct PCRbased detection of new emerging COVID-19 variants.
Sars-cov-2 ; Polymerase Chain Reaction ; Dna Primers ; Oligonucleotide Primers ; Computer Simulation ; Conserved Sequence ; Coronavirus ; Covid-19 ; Disease ; Emergencies ; Evaluation Studies As Topic ; Genes ; Genome ; Global Health ; Health ; Identification (psychology) ; Infection ; Infections ; Membranes ; Methods ; Mutation ; Oligonucleotides ; Organizations ; Population ; Public Health ; Rna ; Rna, Viral ; Sars Virus ; Sequence Alignment ; Severe Acute Respiratory Syndrome ; Syndrome ; Viruses ; Whole Genome Sequencing ; World Health Organization


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