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14 pages, 1544 KB  
Article
Genetic Polymorphisms of IL6-174G/C, TNF-308G/A, and TNF-238G/A and Risk of Pleural Tuberculosis in Venezuelan Patients
by Zaida Araujo, Jacobus Henri de Waard, Mercedes Fernández-Mestre, Douglas Silva, Carmen Judith Serrano, Luis Adrián De Jesús-González, Juan Ernesto Lopez-Ramos and Bruno Rivas-Santiago
Immuno 2026, 6(1), 4; https://doi.org/10.3390/immuno6010004 - 22 Dec 2025
Abstract
Tuberculosis (TB) has various clinical presentations; pulmonary TB (PTB) affects only the lungs, whereas extrapulmonary TB involves other organs, including pleural TB (PLTB). Immunological studies of patients with extrapulmonary TB primarily focus on the cellular Th1 response, which produces key cytokines, including IFN-γ, [...] Read more.
Tuberculosis (TB) has various clinical presentations; pulmonary TB (PTB) affects only the lungs, whereas extrapulmonary TB involves other organs, including pleural TB (PLTB). Immunological studies of patients with extrapulmonary TB primarily focus on the cellular Th1 response, which produces key cytokines, including IFN-γ, TNF, IL-12, and IL-6. TNF and IL-6 play functional roles in host resistance to Mycobacterium tuberculosis (Mtb) infection. Findings suggest that TNF facilitates macrophage containment of Mtb, whereas IL-6 increases macrophage apoptosis induced by Mtb. Studies of the human genome have identified single-nucleotide polymorphisms (SNPs) in genes encoding cytokines associated with TB susceptibility. This study aimed to assess the potential of the IL6-174G/C (rs1800795), TNF-308G/A (rs1800629), and TNF-238G/A (rs361525) SNPs as genetic biomarkers of susceptibility to PLTB in the Venezuelan mestizo population. A total of 269 individuals were included: 69 patients with PLTB and 200 healthy individuals. The IL6-174G/C, TNF-308G/A, and TNF-238G/A polymorphisms were determined by sequence-specific primer polymerase chain reaction (SSP-PCR). Results showed significantly higher frequencies of the G/C, G/A, and G/A genotypes in patients with PLTB (94.0%, 94.2%, and 83.3%) than in controls (40.0%, 19.0%, and 13.4%) for the IL6-174G/C, TNF-308G/A, and TNF-238G/A polymorphisms, respectively. Logistic regression analysis showed significant associations between the G/C, G/A, and G/A genotypes and susceptibility to PLTB. The IL6-174G/C, TNF-308G/A, and TNF-238G/A gene polymorphisms may serve as genetic biomarkers of susceptibility to PLTB in the Venezuelan mestizo population. Full article
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16 pages, 517 KB  
Article
HPV Prevalence and Genotype Distribution Among Infertile and Fertile Women of Turkish Nationality and Association with Cytology and Vaccination Status
by Ayfer Bakır, Büşra Demir Çendek, Muhammed Furkan Kürkçü, Çağlar İzmirli and Murat Aral
Biomedicines 2025, 13(12), 3108; https://doi.org/10.3390/biomedicines13123108 - 17 Dec 2025
Viewed by 121
Abstract
Background/Objectives: Human papillomavirus (HPV) is one of the most common sexually transmitted infections, yet its role in female infertility remains uncertain. This study aimed to compare HPV prevalence and genotype distribution between infertile and fertile women and to evaluate demographic and clinical factors, [...] Read more.
Background/Objectives: Human papillomavirus (HPV) is one of the most common sexually transmitted infections, yet its role in female infertility remains uncertain. This study aimed to compare HPV prevalence and genotype distribution between infertile and fertile women and to evaluate demographic and clinical factors, together with HPV vaccine coverage, in both groups. Methods: Cervical samples from 200 infertile and 200 fertile women aged 18–45 years were analyzed for 28 HPV genotypes using multiplex real-time polymerase chain reaction (PCR). Results: HPV DNA was detected in 13.5% (27/200) of infertile women and 18.0% (36/200) of fertile women (p = 0.272). The most frequent genotypes were HPV-82 (5/200, 2.5%) and HPV-16 (5/200, 2.5%) in infertile women, and HPV-45 (8/200, 4.0%) and HPV-16 (7/200, 3.5%) in fertile women. Single HPV infections were more common in infertile women (81.5%, 22/27) than in fertile women (63.9%, 23/36). HPV positivity was not associated with reproductive, clinical, or lifestyle factors, and age-stratified analyses revealed no statistically significant differences (all p > 0.05). Among women aged 30–45 years, atypical squamous cells of undetermined significance (ASC-US) cytology was identified in eight infertile women, all of whom were HPV-negative, whereas one of nine fertile women with ASC-US was HPV-positive. No low-grade squamous intraepithelial lesion (LSIL) cases were detected in the infertile group. The 9-valent HPV vaccine covered 56.2% (18/32) of genotypes detected in infertile women and 45.1% (23/51) of those detected in fertile women. Conclusions: In this study, no significant differences were observed between the groups in terms of HPV prevalence, genotype distribution, or cytology findings. These results suggest that HPV is not an independent risk factor for infertility and highlight the need for further studies focusing on genotype-specific patterns, viral persistence, and biological mechanisms that may influence reproductive outcomes. Full article
(This article belongs to the Special Issue Current Perspectives on Human Papillomavirus (HPV)—Second Edition)
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18 pages, 2260 KB  
Article
Association Between CYP2C9 and CYP2C19 Genetic Polymorphisms and Antiseizure Medication-Induced Adverse Reactions Among Peruvian Patients with Epilepsy
by Angel T. Alvarado, Felipe L. Ignacio-Cconchoy, Juan C. Espinoza-Retuerto, Roxana M. Contreras-Macazana, Luis Abel Quiñones, Jorge A. García, María R. Bendezú, Haydee Chávez, Felipe Surco-Laos, Doris Laos-Anchante, Pompeyo A. Cuba-Garcia, Elizabeth J. Melgar-Merino, Bertha Pari-Olarte, Mario Bonifaz-Hernández, José Santiago Almeida-Galindo, José Kong-Chirinos, Ricardo Pariona-Llanos, Priscilia Aguilar-Ramírez and Nelson M. Varela
Pharmaceuticals 2025, 18(12), 1872; https://doi.org/10.3390/ph18121872 - 9 Dec 2025
Viewed by 203
Abstract
Background/Objectives: Epilepsy is characterized by recurrent, unprovoked, self-limiting seizures of genetic, acquired, or unknown origin. It affects more than 50 million people worldwide. The prevalence in Peru is 11.9–32.1 per 1000 people. Our objective was to describe the association between CYP2C9 and [...] Read more.
Background/Objectives: Epilepsy is characterized by recurrent, unprovoked, self-limiting seizures of genetic, acquired, or unknown origin. It affects more than 50 million people worldwide. The prevalence in Peru is 11.9–32.1 per 1000 people. Our objective was to describe the association between CYP2C9 and CYP2C19 genetic polymorphisms and adverse reactions induced by antiseizure medications among Peruvian patients with epilepsy. Methods: A descriptive observational study was conducted on Peruvian patients with epilepsy. Non-probability, non-randomized, purposive sampling was carried out through consecutive inclusion. Genomic DNA was obtained from venous blood samples. Genotypes were determined by real-time PCR using specific TaqMan probes to identify the alleles of interest. Results: In total, 89 Peruvian patients with epilepsy were recruited at the Alberto Sabogal Sologuren National Hospital-ESSALUD: 45 were male (23.6 ± 10.0 years) and 44 were female (24.0 ± 12.4 years). The observed frequencies for CYP2C9*2, CYP2C9*3, CYP2C19*2, CYP2C19*3, and CYP2C19*17 were 0.034 (T allele), 0.034 (C allele), 0.14 (A allele), 0.00 (A allele), and 0.03 (T allele), respectively. Patients with intermediate and poor metabolic phenotypes of CYP2C9 and CYP2C19 had a significantly higher risk of adverse drug reactions (ADRs) (OR = 3.75; 95%CI: 1.32–10.69; p = 0.013), compared with normal metabolizers. Polytherapy was a predictor increasing the likelihood of ADRs (OR = 4.33; 95% CI: 1.46–12.80; p = 0.008). Conclusions: In this cohort of Peruvian patients with epilepsy, the reduced-function alleles CYP2C9*2, CYP2C9*3, and CYP2C19*2, associated with decreased metabolic activity, were significantly linked to an increased risk of adverse drug reactions induced by antiseizure medications. Polytherapy further heightened this risk. Collectively, these findings highlight the clinical relevance of CYP2C9 and CYP2C19 genotyping to enhance the safety of antiseizure pharmacotherapy in Latin American settings, where pharmacogenomic evidence remains limited. Full article
(This article belongs to the Section Pharmacology)
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14 pages, 1853 KB  
Article
Identification of Spike Length Gene and Development of KASP Markers in Wheat
by Tiantian Jiang, Lingpeng Meng, Chao Ji, Zehui Wang, Huiwen Cao, Ruoxi Sun, Ke Xu, Xianghai Meng, Xueju Yang and Yong Zhao
Plants 2025, 14(23), 3703; https://doi.org/10.3390/plants14233703 - 4 Dec 2025
Viewed by 307
Abstract
Spike length is a critical trait influencing the yield potential of wheat (Triticum aestivum L.). However, there has been limited research on spike-length-related genes in wheat. Moreover, the scarcity of stable markers for spike-related traits has restricted marker-assisted selection-based breeding. In this [...] Read more.
Spike length is a critical trait influencing the yield potential of wheat (Triticum aestivum L.). However, there has been limited research on spike-length-related genes in wheat. Moreover, the scarcity of stable markers for spike-related traits has restricted marker-assisted selection-based breeding. In this study, a novel long-spike mutant material (LS1) was generated from wheat variety ‘Aikang 58’ (AK58) using ethyl methanesulfonate. We established an F2 segregating population by crossing AK58 with LS1. Morphological analyses of this population indicated that spike length is a dominant quantitative trait regulated by multiple genes. Bulked segregant analysis (BSA) technology was used to preliminarily identify nine candidate regions associated with spike length traits. These regions were mainly in a 7.22 Mb interval (673.84–713.26 Mb) on chromosome 5A and in a 2.34 Mb interval (714.83–717.69 Mb) on chromosome 7B. Twelve candidate genes were identified within these regions. Furthermore, two kompetitive allele specific polymerase chain reaction (KASP) markers (KASP-LS1-681460621 and KASP-LS1-692013966) associated with spike length traits were developed. Both KASP markers effectively genotyped parental lines and the F2 population. Our study results provide a theoretical foundation for the genetic improvement of spike-length-related traits in wheat. Full article
(This article belongs to the Special Issue Cereal Crop Breeding, 2nd Edition)
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16 pages, 3252 KB  
Article
Generation of WW Superfemale Sturgeons Through Hormonal Masculinization of ZW Females
by Ryohei Surugaya, Kazuki Tousaka, Shun Yoshida, Shinji Adachi and Shigeho Ijiri
Fishes 2025, 10(12), 618; https://doi.org/10.3390/fishes10120618 - 2 Dec 2025
Viewed by 225
Abstract
In sturgeon aquaculture, all-female production is desirable due to the high value of caviar. Genetic sexing and the production of WW superfemales are important steps toward achieving this. In this study, we identified the WSR and ZSR primers for amplification of W- and [...] Read more.
In sturgeon aquaculture, all-female production is desirable due to the high value of caviar. Genetic sexing and the production of WW superfemales are important steps toward achieving this. In this study, we identified the WSR and ZSR primers for amplification of W- and Z-specific regions, respectively. WSR primers were designed on the gene W-linked RT RNase H-like domain containing protein (rnhW). The polymerase chain reaction (PCR) bands were obtained with the WSR primer only in phenotypic female sturgeons, indicating that stable genetic sexing was achieved in most species, including those captured around Hokkaido. Moreover, rnhW showed female-specific expression in the gonads during early sex differentiation in kaluga and Amur sturgeon. ZSR primers were developed from the orofacial cleft 1 candidate gene 1 protein homolog. Clear and distinct gel band patterns for ZZ, ZW, and WW genotypes were obtained using WSR and ZSR primers, consistent with genotypic estimations by quantitative PCR. This consistency confirmed the presence of WW superfemales among offspring produced by fertilizing ZW females with ZW pseudomales masculinized using 17α-methyltestosterone. Our findings provide new insights into the mechanisms of sex determination and differentiation in sturgeons, bringing the establishment of an all-female production system within reach. Full article
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19 pages, 823 KB  
Article
Antimicrobial Resistance Phenotypes and Genotypes of Escherichia coli Isolates from Artisanal Minas Frescal Cheeses from the Federal District, Brazil
by Letícia Fernandes Silva Rodrigues, Rodrigo Araújo de Melo, Nathalia Mateus Borges, Anna Cléa Silva Aragão, Marta Oliveira de Araújo, Rebeca Dias dos Santos, Carla Azevedo Bilac, Karolina Oliveira Gomes, Bruno Alcântara do Prado, Lívia Cristina Lira de Sá Barreto, Izabel Cristina Rodrigues da Silva and Daniela Castilho Orsi
Antibiotics 2025, 14(11), 1101; https://doi.org/10.3390/antibiotics14111101 - 2 Nov 2025
Viewed by 785
Abstract
Background/Objectives: This study characterized the phenotypic and genotypic profiles of antimicrobial resistance in 104 Escherichia coli isolates obtained from 22 samples of artisanal Minas Frescal cheese from the Federal District, Brazil. Methods: The antimicrobial susceptibility of E. coli isolates was assessed using [...] Read more.
Background/Objectives: This study characterized the phenotypic and genotypic profiles of antimicrobial resistance in 104 Escherichia coli isolates obtained from 22 samples of artisanal Minas Frescal cheese from the Federal District, Brazil. Methods: The antimicrobial susceptibility of E. coli isolates was assessed using the disk diffusion method and antimicrobial resistance genes were detected using polymerase chain reaction methods with specific primers. Results: The highest rates of phenotypic antimicrobial resistance were observed for sulfonamides (85.58%, 89/104) and tetracyclines (38.46%, 40/104). In the genotypic profiles, most E. coli isolates carried the sulfonamide resistance genes sul1/sul2 (62.50%, 65/104), tetracycline resistance genes tetA/tetB (65.38%, 68/104), and β-lactam resistance genes blaCTX-M/blaTEM/blaSHV (55.77%, 58/104). Most E. coli strains that presented sulfonamide resistance genes carried the sul1 gene (49.04%, 51/104) and were phenotypically sulfonamide-resistant strains (59.61%, 62/104). Regarding the E. coli strains that carried tetracycline resistance genes, the majority harbored both tetA and tetB genes (34.61%, 36/104), with 35.56% (37/104) being phenotypically resistant and 29.80% (31/104) being phenotypically susceptible. For E. coli strains that presented β-lactam resistance genes, the most frequently detected gene was blaCTX-M (21.15%, 22/104) and, notably, most E. coli strains (43.26%, 45/104) were phenotypically susceptible. The cat1 and clmA genes (associated with phenicol resistance) were detected in 22.12% of the E. coli isolates (23/104), with only two strains (1.92%) being phenotypically resistant to chloramphenicol. Conclusion: The high prevalence of E. coli carrying antimicrobial resistance genes in artisanal cheese raises public health concerns regarding the dissemination of potentially pathogenic antimicrobial-resistant microorganisms through the food chain. Full article
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12 pages, 712 KB  
Article
Diagnostic Performance of a Silver-Amplified vs. a Non-Amplified Lateral Flow Kit for Adenoviral Conjunctivitis: A Multicenter Prospective Study
by Tsuguto Fujimoto, Nozomu Hanaoka, Kenichiro Takahashi, Hisatoshi Kaneko, Masaaki Kobayashi, Hisashi Nakagawa, Hiroshi Hatano, Tomoko Tsukahara-Kawamura, Hironori Migita, Kentaro Nakamura, Kiyoharu Kuramoto and Eiichi Uchio
Viruses 2025, 17(11), 1442; https://doi.org/10.3390/v17111442 - 29 Oct 2025
Viewed by 547
Abstract
Accurate diagnosis of adenoviral conjunctivitis is critical for timely treatment and infection control. However, conventional lateral flow kits often lack sufficient sensitivity, especially in mild cases. This multicenter prospective study evaluated the diagnostic performance of silver-amplified lateral flow kits (SA-LFKs) compared with non-silver-amplified [...] Read more.
Accurate diagnosis of adenoviral conjunctivitis is critical for timely treatment and infection control. However, conventional lateral flow kits often lack sufficient sensitivity, especially in mild cases. This multicenter prospective study evaluated the diagnostic performance of silver-amplified lateral flow kits (SA-LFKs) compared with non-silver-amplified kits (NSA-LFKs), using real-time polymerase chain reaction (qPCR) as the reference standard. Tear samples from 200 patients with suspected adenoviral conjunctivitis were collected across four clinics and analyzed for sensitivity, specificity, and genotype coverage. The SA-LFK demonstrated significantly higher sensitivity (86.0%) than the NSA-LFK (72.0%) (p < 0.001), with both kits showing high specificity (100% and 98.1%, respectively). Pooled analysis revealed that sensitivity was significantly lower in mild-to-moderate cases than in severe cases for both kits, suggesting that clinical severity influences detection performance. Across all severity levels, the SA-LFK consistently demonstrated higher sensitivity than the NSA-LFK, including in mild-to-moderate cases (77.8% vs. 59.3%, p = 0.004), supporting its superior diagnostic performance. The SA-LFK showed robust performance across eight identified adenovirus genotypes and maintained higher positivity rates even at lower viral loads. These findings support the clinical utility of SA-LFKs for early diagnosis and outbreak containment in diverse settings. Full article
(This article belongs to the Special Issue Research and Clinical Application of Adenovirus (AdV), 3rd Edition)
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12 pages, 898 KB  
Article
Population-Based Genetic Assessment of Thrombophilia Polymorphisms: Allelic Frequencies and Population Linkage Dynamics
by Panagiota Tsiatsiou, Sofia Balaska, Zafeirios Tsinaris, Maria Archonti, Antonia Lanta, Vasiliki Tsaireli, Anna Takardaki, Areti Kourti, Angeliki Kassomenaki, Georgios Meletis, Dimitrios A. Tsakiris and Lemonia Skoura
Medicina 2025, 61(11), 1935; https://doi.org/10.3390/medicina61111935 - 29 Oct 2025
Viewed by 569
Abstract
Background and Objectives: Thrombophilia is a prothrombotic disorder that increases the risk of blood clotting and can pose serious health problems. It is considered a condition of gene–gene or gene–environment interactions. Variation in the prevalence of thrombophilia mutations and their interaction among [...] Read more.
Background and Objectives: Thrombophilia is a prothrombotic disorder that increases the risk of blood clotting and can pose serious health problems. It is considered a condition of gene–gene or gene–environment interactions. Variation in the prevalence of thrombophilia mutations and their interaction among populations necessitates localized genetic assessments. However, population-based genetic data remains limited for developing effective preventive strategies. Materials and Methods: This cross-sectional observational study was conducted over five years (2020–2024) at a tertiary university hospital in Northern Greece. A total of 2961 individuals aged 18–85 years (mean: 50.5) were registered based on family or medical history of venous thromboembolism (VTE) or clinical symptoms of VTE. The final analysis included 2078 participants comprising 1143 males (55%) and 935 females (45%), who met all the inclusion criteria. Inclusion criteria were absence of acute illness or malignancy, informed consent, and an adequate DNA quantity for genotyping, whereas excluded criteria included incomplete laboratory data, active inflammatory or malignant disease, and cognitive or psychiatric conditions. Peripheral blood samples were collected in 2 mL K3-EDTA tubes, and genomic DNA was analyzed using real-time polymerase chain reaction (PCR) with melting curve analysis and hybridization probes (LightMix® in vitro diagnostics, TIB MolBiol, Berlin, Germany). Five thrombophilia-related polymorphisms, Factor V Leiden (F5 G1691A), prothrombin (F2 G20210A), methylenetetrahydrofolate reductase (MTHFR C677T and MTHFR A1298C), and Plasminogen Activator Inhibitor-1 (PAI-1) 4G/5G, were examined for allele and genotype frequencies, Hardy–Weinberg equilibrium testing, pairwise linkage disequilibrium (D′ and r2), and power analysis. For subjects tested for Factor V Leiden (n = 1476), the activated protein C resistance (APC) ratio was additionally evaluated using the ACL TOP 750 analyzer. Results: Allele frequencies were 7.3% for FV Leiden and 3.7% for FII. The PAI-1 allele was distributed at 44%, while the MTHFR (C677T and A1298C) alleles were each present at 33%. Significant linkage disequilibrium was identified between MTHFR (C677T and A1298C) and between MTHFR A1298C and PAI-1. No evolutionary pressure or demographic bias was found in the Hardy–Weinberg equilibrium. The APC ratio demonstrated a high sensitivity (99.2%) and specificity (96.6%), indicating that it may serve as a reliable screening method. Conclusions: Our findings highlight informative patterns in the genetic predisposition to thrombophilia, which may help develop rule-based strategies for implementing thromboprophylaxis guidelines and personalized medical interventions. Full article
(This article belongs to the Special Issue Genetics of Human Diseases: Fishing for Causality)
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22 pages, 1725 KB  
Article
Exploration of Novel Extracellular Xylanase-Producing Lactic Acid Bacteria from Plant Sources
by Noor Lutphy Ali, Hooi Ling Foo, Norhayati Ramli, Murni Halim and Karkaz M. Thalij
Catalysts 2025, 15(10), 990; https://doi.org/10.3390/catal15100990 - 16 Oct 2025
Viewed by 972
Abstract
Xylanases play a crucial role in bio-transforming sustainable agricultural polymers into xylose-based oligosaccharides, which have great potential in various biotechnology applications. Nevertheless, the application of bacterial xylanase is hindered by the high cost of developing recombinant bacteria to overcome the low activity and [...] Read more.
Xylanases play a crucial role in bio-transforming sustainable agricultural polymers into xylose-based oligosaccharides, which have great potential in various biotechnology applications. Nevertheless, the application of bacterial xylanase is hindered by the high cost of developing recombinant bacteria to overcome the low activity and narrow pH stability. Considerable efforts have been made to discover and explore new wild bacterial strains that produce highly effective and environmentally sustainable extracellular xylanase enzymes for various targeted biotechnological and industrial applications. Lactic acid bacteria (LAB) have recently been proven to be versatile producers of extracellular hydrolytic enzymes. Therefore, this study aimed to isolate and characterise extracellular xylanase-producing LAB (EXLAB) from plant sources. The specific extracellular xylanase activity was determined across a wide pH range, from acidic to alkaline. Subsequently, the expression of xylanase genes of EXLAB grown under acidic and alkaline conditions was determined by quantitative reverse transcription polymerase chain reaction. A total of 45 putative LAB were isolated from radish, gundelia and rhubarb plants. They were identified by phenotypic and genotypic approaches. However, only 15 LAB isolates were confirmed as EXLAB. Weissella confusa and Pediococcus pentosaceus were the most common species among the identified EXLAB. The XylW (~196 bp) and XylP (189 bp) xylanase genes were then amplified from W. confusa and P. pentosaceus, respectively. P. pentosaceus G4 demonstrated the most versatile extracellular xylanase production that was active from pH 5 to pH 8. However, a significant increase in extracellular xylanase gene expression (13.45-fold) at pH 5 was noted as compared to pH 8. Similarly, P. pentosaceus G4 also exhibited the highest extracellular xylanase activity (0.88 U/mg) at pH 5. This study reveals the potential of P. pentosaceus G4 as an eco-friendly and novel extracellular xylanase producer possessing broad pH stability. The robust gene expression and activity of extracellular xylanase imply P. pentosaceus G4 is a promising candidate for sustainable enzymatic processes essential for the environmentally friendly enzymatic reactions and applications. Full article
(This article belongs to the Section Biocatalysis)
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15 pages, 1187 KB  
Review
Integration of Point-of-Care Technology in the Decoding Process of Single Nucleotide Polymorphism for Healthcare Application
by Thi Ngoc Diep Trinh, Hanh An Nguyen, Nguyen Pham Anh Thi, Thi Xuan Tuy Ho, Kieu The Loan Trinh and Nguyen Khoi Song Tran
Micromachines 2025, 16(10), 1159; https://doi.org/10.3390/mi16101159 - 13 Oct 2025
Cited by 1 | Viewed by 935
Abstract
Single nucleotide polymorphism (SNP) involves plenty of genetic disorders in organisms that can be passed down to the next generation or cause the stimulant signal that leads to early mortality in infants, especially within humankind. In medical field, real-time polymerase chain reaction (RT-PCR) [...] Read more.
Single nucleotide polymorphism (SNP) involves plenty of genetic disorders in organisms that can be passed down to the next generation or cause the stimulant signal that leads to early mortality in infants, especially within humankind. In medical field, real-time polymerase chain reaction (RT-PCR) is the most popular method for disease diagnosis. The investigation of genetic maps for the prediction of inherited illnesses needs the collaboration of sequencing technique and genome analysis. Although these methods are popular now, the cost for each test is quite high. Moreover, there is the requirement of extra machines and skillful technician or specialist level. Among these popular methods, the allele-specific polymerase chain reaction (AS-PCR), allele-specific loop isothermal mediated amplification (AS-LAMP), and allele-specific recombinase polymerase amplification (AS-RPA) are brought up for screening the nucleotide differences in the genetic sequence which will be noticed in this review as their availability, novelty, and potential for quick distinguishing of disease caused by SNP. Point-of-care testing (POCT) is a system built in a portable size but can perform the entire process of SNP recognition. Along with that, the POCT is intersected with the mentioned amplification methods and the genetic material preparation steps to become a united framework for higher efficiency and accuracy and lower cost. According to that, this review will focus on three common amplification techniques and their combination with POCT in the upstream and downstream process to genotype SNP related to human diseases. Full article
(This article belongs to the Section B4: Point-of-Care Devices)
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19 pages, 6041 KB  
Article
Integrating RPA-LFD and TaqMan qPCR for Rapid On-Site Screening and Accurate Laboratory Identification of Coilia brachygnathus and Coilia nasus in the Yangtze River
by Yu Lin, Suyan Wang, Min Zhang, Na Wang, Hongli Jing, Jizhou Lv and Shaoqiang Wu
Foods 2025, 14(20), 3484; https://doi.org/10.3390/foods14203484 - 13 Oct 2025
Viewed by 572
Abstract
Accurate differentiation between Coilia brachygnathus and Coilia nasus is imperative for the effective management of fisheries, the conservation of aquatic ecosystems, and the mitigation of commercial fraud. Current morphological identification remains challenging due to their high morphological similarity—particularly for processed samples—while conventional molecular [...] Read more.
Accurate differentiation between Coilia brachygnathus and Coilia nasus is imperative for the effective management of fisheries, the conservation of aquatic ecosystems, and the mitigation of commercial fraud. Current morphological identification remains challenging due to their high morphological similarity—particularly for processed samples—while conventional molecular methods often lack the speed or specificity required for field applications or high-throughput screening. In this study, a novel integrated approach was developed and validated, combining TaqMan quantitative real-time PCR (qPCR). for precise genotyping of C. brachygnathus and C. nasus with Recombinase Polymerase Amplification coupled with Lateral Flow Dipstick (RPA-LFD) for rapid on-site screening. First, species-specific RPA-LFD assays were designed to target the mitochondrial COI gene sequence. This enabled visual detection within 10 min at 37 °C, with a sensitivity of 102 copies/μL, and required no complex equipment. A dual TaqMan MGB qPCR assay was further developed by validating stable differentiating SNPs (chr21:3798155, C/T) between C. brachygnathus and C. nasus, using FAM/VIC dual-labeled MGB probes. Results showed that this assay could distinguish the two species in a single tube: for C. brachygnathus, Ct values in the FAM channel were significantly earlier than those in the VIC channel (ΔCt ≥ 1), with a FAM detection limit of 125 copies/reaction; for C. nasus, only VIC channel amplification was observed, with a detection limit as low as 12.5 copies/reaction. Validation with 171 known tissue samples demonstrated 100% concordance with expected species identities. This integrated approach effectively combines the high accuracy and quantitative capacity of TaqMan qPCR for confirmatory laboratory genotyping with the speed, simplicity, and portability of RPA-LFD for initial field or point-of-need screening. This reliable, efficient, and user-friendly technique provides a powerful tool for resource management, biodiversity monitoring, and ensuring the authenticity of high-quality C. brachygnathus and C. nasus. Full article
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24 pages, 4004 KB  
Article
Genetic Monitoring of the Endangered Acipenser dabryanus Using a High-Resolution MNP System
by Lu Cai, Wei Jiang, Zhiwei Fang, Hai Peng, Hao Chen, Renjing Wan, Lifen Gao, Baolong Zhang, Zilan Xiao, Sha Li, Lun Li, Lihong Chen, Huiyin Song, Tiantian Li and Junfei Zhou
Diversity 2025, 17(10), 704; https://doi.org/10.3390/d17100704 - 11 Oct 2025
Viewed by 548
Abstract
Acipenser dabryanus, once abundant in China’s freshwater ecosystems, is now extinct in the wild. Effective genetic tools are urgently needed to support conservation efforts under the Yangtze River Protection Law and the 10-year fishing ban. Traditional molecular markers (e.g., COI, SSR, [...] Read more.
Acipenser dabryanus, once abundant in China’s freshwater ecosystems, is now extinct in the wild. Effective genetic tools are urgently needed to support conservation efforts under the Yangtze River Protection Law and the 10-year fishing ban. Traditional molecular markers (e.g., COI, SSR, SNP) often lack sufficient resolution for fine-scale population assessment. Here, we developed a high-resolution Multiple-Nucleotide Polymorphism (MNP) system for A. dabryanus, comprising 424 newly developed, highly polymorphic markers optimized for multiplex PCR and high-throughput sequencing. The MNP system demonstrated excellent performance in individual fin tissue samples, successfully distinguishing Acipenser sinensis and Acipenser ruthenus individuals from the A. dabryanus population. In addition, 41 characteristic alleles specific to A. dabryanus were further identified. Across samples, it achieved >90% MNP locus detection rate, with an average of 7.48 alleles per locus, 66.5% heterozygosity, >98% reproducibility, and 99% accuracy. A strong correlation was observed between DNA concentration and spike-in-based copy numbers (R2 > 0.99), and sensitivity analysis confirmed reliable detection at ~1 copy/reaction. Application of the system across 97 samples, including 51 A. dabryanus tissue samples and 46 water environmental samples, revealed clear population structure with an average genetic differentiation of 70.45%, highlighting substantial genetic diversity within the sampled populations. Based on the above experimental results, the high-resolution MNP system has the potential to enable construction of population-specific allelic genotypes to distinguish wild individuals from released ones and, when applied to tissue and eDNA samples, to facilitate monitoring of migration pathways and habitat connectivity. Such applications could provide essential genetic information to evaluate release programs, guide conservation strategies, and inform habitat restoration for the recovery of A. dabryanus. Full article
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15 pages, 1267 KB  
Article
Genetic Variations of the FUT3 Gene in Le(a−b−) Individuals and Their Association with Lewis Antibody Responses
by Oytip Nathalang, Piyathida Khumsuk, Wiradee Sasikarn and Kamphon Intharanut
Med. Sci. 2025, 13(4), 218; https://doi.org/10.3390/medsci13040218 - 2 Oct 2025
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Abstract
Background: The biosynthesis of Lewis (Le) antigens depends on the FUT3 gene, encoding an α(1,3/4)-fucosyltransferase. Individuals lacking functional FUT3 exhibit a Le(a–b–) phenotype, regardless of secretor status. Methods: This study determined the prevalence of FUT3 single nucleotide variants (SNVs) in Thai blood donors [...] Read more.
Background: The biosynthesis of Lewis (Le) antigens depends on the FUT3 gene, encoding an α(1,3/4)-fucosyltransferase. Individuals lacking functional FUT3 exhibit a Le(a–b–) phenotype, regardless of secretor status. Methods: This study determined the prevalence of FUT3 single nucleotide variants (SNVs) in Thai blood donors and characterised genotype and allele distributions. We also examined the association between FUT3 variants and the presence of Le antibodies to better understand variability in immune responses. A total of 112 blood donors were recruited, comprising 52 non-responders and 60 responders for Le antibody detection. The FUT3 coding sequence was amplified by polymerase chain reaction and directly sequenced to identify single nucleotide variants (SNVs) and haplotypes. Results: Associations between FUT3 SNVs, haplotypes, and Le antibody responsiveness were subsequently analysed. Thirteen FUT3 SNVs were identified, with c.59T>G (rs28362459) present in all Le(a–b–) cases. The FUT3*01N.17.03 (le59,1067) haplotype was most common (0.634) and showed the strongest association with Le antibody responsiveness (adjusted OR = 3.052, 95% CI: 1.683–5.534, p < 0.0001). Differences in antibody types, isotypes, and the FUT3*01N.17.03 genotype between groups were not statistically significant. Conclusions: This first study characterises FUT3 variations in Le(a–b–) Thai blood donors and identifies FUT3*01N.17.03 as associated with Le antibody responsiveness, highlighting its relevance for population-specific genetic diagnostics in transfusion medicine. Full article
(This article belongs to the Section Translational Medicine)
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12 pages, 865 KB  
Article
Genetic and Clinical Insights into ALS/FTD: Profiling a Rare Cohort to Explore Spectrum Heterogeneity
by Ana Marjanovic, Elka Stefanova, Vanja Viric, Aleksa Palibrk, Gorana Mandić Stojmenović, Tanja Stojković, Lenka Stojadinovic, Ivana Basta, Ivana Novakovic, Zorica Stević and Milena Jankovic
J. Pers. Med. 2025, 15(10), 451; https://doi.org/10.3390/jpm15100451 - 28 Sep 2025
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Abstract
Background: Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are recognized as a spectrum of neurodegenerative disorders with overlapping clinical, pathological, and genetic features. The identification of C9orf72 hexanucleotide repeat expansion as the most common genetic cause of both conditions has prompted [...] Read more.
Background: Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are recognized as a spectrum of neurodegenerative disorders with overlapping clinical, pathological, and genetic features. The identification of C9orf72 hexanucleotide repeat expansion as the most common genetic cause of both conditions has prompted further investigation of genetic modifiers that may contribute to disease heterogeneity. We aimed to analyze the frequency of C9orf72 repeat expansions and potential modifying roles of APOE, ATXN1, and ATXN2 in Serbian ALS/FTD patients. Methods: Our study included an ALS/FTD cohort (n = 22) and healthy controls (n = 94). Repeat sizing in C9orf72, ATXN1 and ATXN2 was performed by fluorescent polymerase chain reaction (PCR) and capillary electrophoresis, while repeat-primed PCR was used to confirm C9orf72 expansions. APOE genotyping was conducted using real-time PCR assays targeting SNPs rs429358 and rs7412. Results: In the ALS/FTD cohort, 31.82% of the patients had heterozygous C9orf72 repeat expansion. The most common APOE genotype among patients was ε3/ε3 (72.73%). Intermediate-length ATXN1 alleles (32–44 repeats) were detected in 13.64% of patients and ATXN2 intermediate-length alleles (27–33 repeats) were found in 9% of patients. No significant differences were observed between ALS/FTD patients and controls in APOE ε4 frequency or intermediate ATXN1/ATXN2 repeats. Conclusions: Larger, population-specific studies and meta-analyses are needed to better understand the role of genetic modifiers in ALS/FTD pathogenesis and their influence on clinical heterogeneity. By integrating genetic and clinical data, this study represents a step toward the development of precision medicine strategies for ALS/FTD. Full article
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16 pages, 1580 KB  
Article
Polymorphism of the BMPR1B Variants for Prolific Traits in the Indonesian Local Ettawah Goat
by Mudawamah Mudawamah, Muhammad Zainul Fadli, Gatot Ciptadi, Fatchiyah Fatchiyah, Mahayu Woro Lestari, Yudith Oktanella, Susiati Susiati and Albert Linton Charles
Animals 2025, 15(19), 2781; https://doi.org/10.3390/ani15192781 - 24 Sep 2025
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Abstract
The Indonesian Local Ettawah Goat (ILEG) exhibits substantial genetic variation, suggesting its potential for high productivity and promote sustainable practices in farm animal breeding. This study aimed to investigate the molecular characteristics of prolific ILEG by identifying potential candidate genes through polymerase chain [...] Read more.
The Indonesian Local Ettawah Goat (ILEG) exhibits substantial genetic variation, suggesting its potential for high productivity and promote sustainable practices in farm animal breeding. This study aimed to investigate the molecular characteristics of prolific ILEG by identifying potential candidate genes through polymerase chain reaction (PCR) analysis of the bone morphogenetic protein receptor type 1B (BMPR1B) gene with two variants: alleles G and A. The research involved PCR amplification and sequencing of the BMPR1B A allele, followed by a combined PCR approach integrating both A and G alleles for genotyping. Blood samples were collected from 73 does with documented prolificacy history and 358 does without prolificacy histories, sourced from seven village breeding operations in East Java. PCR amplification yielded fragments of 556–1181 base pairs in all samples. Haplotype analysis revealed 15 unique haplotypes with a diversity of 0.94 and a mutation frequency of 27.15%. Integration of the BMPR1B alleles G and A revealed polymorphic prolific traits. Polymorphism analysis of 385 ILEGs demonstrated allele frequencies of 0.55 for allele A and 0.45 for the allele G. Average fecundity rates associated with the BMPR1B polymorphism were 1.49 offspring for the homozygous AA, 1.60 for the heterozygous GA, and 1.89 for the homozygous GG. While overall differences among genetic groups were approached statistically significantly (Kruskal–Wallis, p = 0.056), pairwise comparison (Mann–Whitney test) revealed that homozygous GG was significantly associated with higher prolificacy compare to the heterozygous GA (p = 0.029) and homozygous AA (p = 0.040). Similar results were also obtained from data without documented history. These findings suggest that the GG polymorphism of BMPR1B may increase prolificacy in ILEG. Furthermore, the higher frequency of allele G highlights the importance of considering prolificacy traits in breeding selection strategies to enhance sustainable genetic improvement and increase litter size in ILEG. It is recommended to apply dual-primer specific amplification and fragment size differentiation as key molecular approaches for polymorphism of the BMPR1B gene and prolificacy, since these methods can highlight genetic variation and provide valuable markers for breeding programs of the Indonesian Local Etawah Goat. Full article
(This article belongs to the Section Small Ruminants)
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