Accelerating quantitative polymerase chain reaction (qPCR) without compromising analytical fidelity remains a significant challenge in molecular diagnostics, primarily due to the thermal lag between heating elements and reagents. Here, we report a high-speed qPCR platform that overcomes this limitation using a reagent-centric cascade control strategy. The system employs a planar PCB-based copper heater that functions as both a heating element and a temperature sensor, ensuring low-latency, sensor-efficient thermal feedback. To overcome this thermal delay, a virtual temperature sensor-derived from system identification-is used to estimate the real-time reagent temperature, which drives an outer-loop fuzzy PID controller with feedforward compensation. Inner cascaded loops stabilize the heater current and surface temperature. The system achieves reagent-phase average heating and cooling rates of 24.1 °C s-1 and 19 °C s-1, respectively. Furthermore, the reagent temperature is controlled with an accuracy of ±0.2 °C and an overshoot of less than 0.2 °C. A complete 45-cycle amplification is achieved in as little as 4.4 minutes. Crucially, this speed is attained without analytical compromise, demonstrating excellent quantitative accuracy (R2 = 0.9965) and amplification efficiency (109.8%). The proposed reagent-centred control framework offers a scalable pathway for developing high-throughput PCR and molecular diagnostic instruments, supporting fast, accurate, and scalable nucleic acid testing.

Health Economic Benefits of Implementing Nucleic Acid Testing in Addition To Serology for Blood Transfusion Safety in India: a Cost-Effectiveness Analysis

Dehydroepiandrosterone opposes cardiac aging via NFκB/IL-10/Sirt1/Nrf2 mediated pathway in aged rats.

HIV screening in blood donation: Diagnostic accuracy of chemiluminescence and rapid test compared to nucleic acid test in a large-scale Eastern India cohort.

A quick thermal response digital acoustofluidic system for rapid on-chip PCR.

Biallelic ACSF3 variants with combined malonic and methylmalonic acidemia and associated developmental epileptic encephalopathy phenotype: A novel genotype-phenotype correlation.

The preventive effect of nirsevimab, a long-acting monoclonal antibody, on infants with respiratory syncytial virus (RSV) infection beyond six months has rarely been evaluated. The objective of this study was to evaluate the preventive effects of nirsevimab beyond 6 months after RSV infection using a global database. This was a multi-centre retrospective study using a global database (TriNetX). The participants were children under 24 months of age who required nucleic acid testing for RSV between July 2023 and June 2025. Children who received the last dose of nirsevimab within 6 months, between 6 and 11 months, and beyond 12 months were compared with those who did not receive any nirsevimab dose by epidemic season after propensity score matching. A total of 4627, 861, and 532 children aged <24 months who received nirsevimab within 6 months, between 6 and 11 months, and beyond 12 months before RSV testing, respectively, and 210,626 children who did not receive any nirsevimab were identified. Compared with those who did not receive nirsevimab, the odds ratios (OR) of a positive RSV test result among those who received nirsevimab within 6 months, between 6 and 11 months, and beyond 12 months before testing were 0.49 [95% confidence interval (CI) 0.42, 0.57] (p<0.001), 0.67 [0.48, 0.94] (p<0.020), and 1.21 [0.89, 1.65] (p=0.234) in the last epidemic season, respectively. This study indicated that the preventive effect of nirsevimab against RSV infection was maintained for up to 12 months following administration, whereas a preventive effect beyond 12 months was not observed. The results of this study need to be interpreted with caution owing to some important limitations.

Human papillomavirus (HPV) is a frequent sexually transmitted infection that can cause cervical cancer in women. Early detection could slow the spread of HPV infection and its eventual development into cervical cancer. Furthermore, there are few statistics available on the prevalence of cervical cancer and HPV in Ethiopia. Therefore, the purpose of this study was to assess the prevalence of high-risk cervical human papillomavirus infections and the factors associated with them among women who visited the family planning and gynecology clinic at Debre Tabor Comprehensive Specialized Hospital in Northcentral Ethiopia. A health facility-based cross-sectional study was conducted among women attending the gynecology and family planning clinic from October 2022 to May, 2023. After obtaining written informed consent from each participant woman, the data on sociodemographic and clinical factors were collected by a trained nurse through face-to-face interviews using a pretested questionnaire. Clinicians collected cervical swabs from participants, which were then analyzed by laboratory technologists using the OncoE6™ Cervical Test (Arbor Vita Corporation, Fremont, CA, USA) a lateral flow assay coated with high-affinity monoclonal antibodies designed to detect E6 oncoproteins specific to high-risk HPV types 16 and 18. In addition, clinicians conducted the Visual Inspection with Acetic Acid (VIA) test, a procedure in which diluted acetic acid is applied to the cervix; areas with abnormal epithelial cells temporarily turn white, allowing for visual identification of potential lesions. The data was analyzed using SPSS version 21, using bivariate and multivariate logistic regression models, with a p value of ≤ 0.05 indicating statistical significance. There were 335 study participants during the course of the study. The mean age of the study participants in this study was 36.66 ± 7.92 years. Furthermore, the overall prevalence of HPV (HPV16 and/or 18) was 13.4%, with a 95% CI of 10.1%-17.3%. Fifty-three individuals (15.8%) out of the total study participants tested positive for the VIA. Moreover, the overall prevalence of HPV type 16, HPV18, and HPV16 and 18 coinfections was 8.66%, 7.76%, and 2.99%, respectively. A previous history of STIs (AOR = 14.42, 95% CI = 6.31-32.99, P = 0.001), HIV infection (AOR = 3.53, 95% CI = 1.46-8.54, P = 0.005) and age between 40 and 49 years (AOR = 0.17, 95% CI = 0.04-0.72, P = 0.016) were significantly associated with human papillomavirus infection. The results of the current study showed a significant prevalence of HPV infection and a high VIA positive rate. This study also demonstrated that age, HIV infection, and a history of STIs were significantly associated predictors of HPV infection. Overall, these findings highlight the importance of early screening, education, and prevention efforts to reduce the prevalence of HPV infections and associated diseases among women in the study area.

Toxoplasma gondii is a zoonotic pathogen that can infect humans and a wide range of warm-blooded animals, posing a significant threat to human health and the livestock industry. The development of a time-saving, highly sensitive, and specific method for the detection of T. gondii in tissue and blood samples is crucial to the monitoring, prevention, and control of toxoplasmosis. In this study, we evaluated the efficiency of a previously described method, termed REPORT, that integrates recombinase polymerase amplification with CRISPR/Cas12a for the detection of T. gondii nucleic acids. We evaluated the limit of detection (LOD) and specificity of the extended REPORT method using prepared target DNA in addition to tissue and blood samples. Furthermore, we validated the accuracy of T. gondii detection in clinical samples using the REPORT-based method in comparison with nested PCR based on the B1 gene. Sensitivity tests showed that the LOD of the REPORT-based fluorescence method and the lateral flow strip method were 3.7 copies /μL for target DNA, 3.1 tachyzoites/g for tissue samples, and five tachyzoites/mL for blood samples. Specificity tests suggested that the REPORT method had good specificity and did not cross-react with several common parasites. The method performed well for clinical DNA samples, demonstrating its ability for use in on-site detection.IMPORTANCEToxoplasma gondii can infect over 200 species of warm-blooded animals, including humans, posing not only a significant threat to public health systems but also causing substantial economic losses to the global livestock industry. Current diagnostic methods are slow, equipment-dependent, and impractical for field use. This study addresses these limitations by developing REPORT, a rapid, ultrasensitive nucleic acid test combining recombinase polymerase amplification and CRISPR/Cas12a. The REPORT detects T. gondii in tissue and blood samples within 1 h at low cost, requiring only a portable heater. Its visual results (fluorescence or test strips) enable on-site use without specialized training, achieving 100% accuracy versus nested PCR. With a sensitivity of 3.1 parasites per gram of tissue and five parasites per milliliter of blood, this method revolutionizes toxoplasmosis screening in resource-limited clinics, farms, and food safety inspections, empowering timely interventions to curb transmission and improve public health outcomes.

Cellulose acetate (CA) is a semi-synthetic polymer widely present in modern and contemporary collections, yet its conservation poses major challenges due to its chemical and physical instability. Hydrolytic degradation, acetic acid release, plasticizer loss, and embrittlement compromise both structure and surface, making cleaning particularly difficult. Conventional cleaning methods may cause abrasion, extract additives, or alter gloss. Although hydrogels have shown promise for CA cleaning, the literature remains extremely limited. This study reports a preliminary investigation of gel-based cleaning on Le Rituel (1968), a heavily soiled cellulose acetate (CA) artwork by José Escada. The object’s condition was assessed through visual inspection, pH measurements, volatile acidity testing, and infrared spectroscopy. Cleaning tests were conducted on a CA replica (2006) with superficial soiling and on selected artwork areas. Two gel formulations were evaluated: the biopolymer agar-agar rigid gel and the synthetic viscoelastic poly(vinyl alcohol)-borax (PVAl-Borax) gel. Agar-agar was effective as a first step, reducing superficial soiling and humidifying adherent residues for subsequent removal, while PVAl-Borax was advantageous in the second step, as its viscoelastic properties enabled controlled mechanical action and facilitated the removal of more adherent residues. This case study demonstrates the potential of combined gel systems as versatile tools for CA conservation.

ObjectiveThis study aimed to compare the diagnostic efficacy between the Kehua and Roche nucleic acid testing (NAT) systems for detecting hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) in blood donation screening.MethodsWe analyzed retrospective data from 670,775 transient blood donation samples collected between 2016 and 2024. Key performance indicators (KPIs) included the types of reactive samples, effective split rate, effective reaction rate, single pool invalid number, batch invalid number, and pooling missing samples. Annual trends in cycle threshold (CT) value distributions and overall system performance were also evaluated.ResultsAmong 419 reactive samples (Kehua: 202; Roche: 217), no significant differences were observed in the effective split rates (P > 0.05) or overall specimen reaction rates (P > 0.05) between the two systems. The HCV detection rate was significantly lower for the Kehua system (P < 0.05), while the detection rates for HBV and HIV showed no significant inter-system differences (P > 0.05). The separation efficiency of reactive pools varied significantly across different CT value ranges (P < 0.05). The Kehua system demonstrated stable annual effective split and reaction rates (P > 0.05), whereas the Roche system exhibited significant annual fluctuations in these metrics (P < 0.05). The Kehua system had significantly fewer single pool invalid numbers and pooling omissions than the Roche system (P < 0.05).ConclusionThe Kehua and Roche NAT systems demonstrated comparable overall performance in blood screening, with Kehua proving non-inferior to Roche. Kehua’s advantages included fewer invalid tests and fewer pooling errors, which could reduce economic and time costs. The Roche system exhibited higher automation, supporting continuous batch processing. The observed CT value-dependent separation efficiency suggests potential for protocol optimization in detecting low viral load samples.

Background: Hepatitis B virus (HBV) remains a transfusion-transmissible infection of global concern. While mandatory screening for hepatitis B surface antigen (HBsAg) has reduced overt infections, occult hepatitis B infection (OBI) poses ongoing risk. This meta-analysis aimed to estimate the pooled prevalence of anti-HBc, HBsAg, and OBI in Saudi blood donors, as well as to assess regional variations and temporal trends. Methods: A systematic meta-analysis was conducted using nine studies published between 2013 and 2024, encompassing a total of 87,820 blood donors. Prevalence was pooled using random-effects models with logit transformation and Hartung–Knapp adjustment. Heterogeneity was quantified with I2 and τ2. Subgroup analyses examined geographic regions; meta-regression assessed publication year. Publication bias was evaluated with Egger’s regression; sensitivity analyses tested robustness. Results: The pooled prevalence of anti-HBc was 5% (95% CI: 3–7%), HBsAg was 0.46% (95% CI: 0.31–0.69%), and OBI was 0.12% (95% CI: 0.03–0.39%). High heterogeneity was observed (I2 = 90.6–98.9%). Subgroup analyses revealed regional disparities, and meta-regression showed no significant temporal trends. Conclusions: The findings suggest that past HBV exposure remains relatively common among Saudi blood donors, while current active infection and OBI are infrequent. These results support continued enhanced screening, including anti-HBc and nucleic acid testing (NAT), to ensure transfusion safety. The lack of a national hemovigilance system that monitors transfusion-transmitted infections among recipients represents a critical gap in Saudi Arabia’s transfusion safety framework. Addressing this gap is essential to fully assess residual risks, evaluate the real-world effectiveness of screening policies, and align with global best practices in blood safety.

Introduction. Transfusion Transmissible Infections (TTIs) remain a major concern in ensuring the blood safety. Nucleic acid testing (NAT) improves detection by shortening the window period (WP) and identifying occult infections. This study aims to determine the significance of NAT through a comparative analysis of the chemiluminescent serological technique (CMIA) and the molecular technique (NAT). Methods. This retrospective study analyzed TTI testing data from 167979 blood donations (93,390 donors) between 2022 and 2024. Screening included HBV DNA, HCV RNA, and HIV RNA (Procleix UltrioPlex E, Panther System) and serological markers (HBsAg, anti-HCV, anti-HIV/p24) using the Architect 2000 platform. Repeatedly seroreactive samples underwent confirmatory testing (HBsAg neutralization, HCV/HIV immunoblot), while NAT-reactive samples were individually tested using the Procleix Ultrio Elite discriminatory assay. Results. Serological testing identified 237 (0.25%) confirmed TTI cases, while NAT detected 245 (0.26%), including 22 (0.023%) NAT-only positive cases. Among these, 21 were HBV DNA-positive/HBsAg-negative. Follow-up samples from 11 donors showed seroconversion in 7 (33.3%), while 4 (19.0%) were identified as potential occult HBV infections (OBI). One HIV NAT-only case was detected. The NAT yield was 1 per 4,245 donations, decreasing over time. Residual risk per 100,000 donations declined from 10.55 to 4.59 for HBV, while remaining low for HCV and HIV (0.12 in 2024). Conclusion. NAT enhances blood safety by detecting early and occult infections missed by serology. The declining NAT yield and residual risk emphasize improved screening efficiency. Integrating NAT with serological methods is crucial for reducing transfusion-transmitted infections.

BackgroundBased on China’s current HIV-1 testing algorithm, samples with initial positive antibody screening require two additional repeat tests, with infection status confirmed by HIV-1 antibody Western Blot or nucleic acid testing (NAT). In recent years, NAT has gradually become the mainstream supplemental assay due to its shorter window period, holding significant importance in detecting acute HIV-1 infection. Currently, over 800 laboratories in China can perform HIV-1 NAT. However, most NAT kits are designed for viral load (VL) detection, typically used to assess treatment efficacy in confirmed patients. This study aimed to explore the diagnostic efficacy of HIV-1 quantitative NAT at varying medical decision points, as well as the methodological preference for antibody screening, to support better implementation of HIV-1 testing and accurate identification of HIV-1 infections in China.MethodsTesting records of 11,369 samples with WB-indeterminate or -negative results during 2018–2023 were collected from the central HIV confirmatory laboratory databases of 20 provincial CDCs. Methodological preferences in HIV-1 antibody screening, diagnostic performance of single- or double-reactive results in retesting and quantitative NAT at varying diagnostic thresholds were analyzed.ResultsChemiluminescence immunoassay (CLIA) was the most widely used method for initial screening (42.77%) and the seroconversion rate was significantly higher in cases with double-positive retest results compared to single-positive results (75.95% vs. 27.25% for WB-indeteminate results and 20.31% vs. 7.26% for WB-negative results, respectively). The specificity and positive predictive values of HIV-1 quantitative NAT were 100% at each medical decision points, and the sensitivity and negative predictive value decreased from 99.92% to 99.93% to 94.71% and 95.64%. Two samples with undetectable viral loads were ultimately diagnosed as HIV-1 infections, alerting us to consider individual exposure history when managing antibody screening-reactive but VL-undetectable cases.ConclusionsQuantitative HIV-1 NAT enables rapid and accurate infection confirmation, earlier identification and timely treatment initiation.

A universal, dual-mode microfluidic platform is presented for high-throughput (HTP) biochemical analysis, capable of performing either enzyme-linked immunosorbent assays (ELISA) as well as rapid nucleic acid (NA) purification within a single lab-on-a-disc. Assay-specific functionality is achieved by simply coating antibodies for immunoassays or inserting glass fiber membranes for NA extraction, eliminating the need for multiple chip designs. A self-pressure-balanced zigzag distributor generates a capillary pressure gradient that enables fast and uniform reagent distribution across all 30 units with a single injection, preventing overflow and residual blockage under high-speed injection or repeated centrifugation steps. In addition, integrated siphon valves and bifurcation chambers precisely control fluid movement for incubation, washing, detection, and elution steps. In ELISA mode, the platform automates the full workflow and delivers results for 30 tests in under 1h. In NA mode, RNA is extracted and eluted for downstream qPCR analysis. Using SARS-CoV-2 as a model, the platform demonstrates the ability to detect nucleocapsid protein via ELISA and confirm results through qPCR, enabling orthogonal validation and enhanced diagnostic reliability. This reconfigurable, dual-mode system offers a unified solution for protein and nucleic acid testing, ideally suited for scalable deployment in clinical diagnostics, outbreak response, and translational research.

Climate change and air pollution have significantly influenced disease prevalence and transmission. Human adenovirus (HAdV) is a common pathogen that causes acute respiratory infections in children. This study employed an ecological approach to investigate the relationship between HAdV infection in children in Lanzhou, Northwest China, and meteorological factors and air pollutants, aiming to deepen the understanding of environmental factors affecting viral transmission. Clinical specimens from children with acute respiratory infections were collected at a sentinel hospital in Lanzhou from January 1, 2023, to February 28, 2025, and were subjected to respiratory adenovirus nucleic acid testing. HAdV infection patterns were analyzed according to age, sex, and season. A combined approach of stepwise linear regression and generalized additive models (GAM) was employed to investigate the correlations between HAdV infection and meteorological factors/air pollutants from January 1, 2023, to December 31, 2024. Between January 1, 2023, and February 28, 2025, a total of 1,339 throat swab specimens were collected from children with acute respiratory infections (ARIs), with a male-to-female ratio of 1.48:1. The HAdV incidence rate in 2024 (11.23%) was higher than that in 2023 (5.45%). The HAdV incidence rates among male and female children were 5.15% and 3.73%, respectively. Preschool-and school-aged children exhibited higher incidence rates than infants and toddlers, at 3.29%, 3.36%, 0.75%, and 1.49%, respectively. HAdV circulated throughout all four seasons, with peak prevalence in autumn (2.28%) and winter (5.06%). HAdV incidence rates showed significant negative correlations with meteorological factors (air temperature, sunshine duration, wind speed), with correlation coefficients of: r = -0.6398 (95% CI: -0.8878, -0.1042), (P = 0.0250); r=-0.6376 (95% CI: -0.8870, -0.1005), (P = 0.0257); r=-0.6208 (95% CI: -0.8809, -0.07285), (P = 0.0312); showed significant positive correlations with atmospheric pollutants (CO, NO₂, and SO₂), with correlation coefficients of: r = 0.7610 (95% CI: 0.3321, 0.9291), (P = 0.0040); r = 0.6846 (95% CI: 0.1824, 0.9035), (P = 0.0140); r = 0.7162 (95% CI: 0.2147, 0.9143), (P = 0.0088); while exhibiting a significant negative correlation with atmospheric pollutant O₃, with a correlation coefficient of r = -0.6938 (95% CI: -0.9067, -0.1992), (P = 0.0123). GAM analysis revealed that meteorological factors (air temperature, sunshine duration, and wind speed) were all nonlinearly associated with adenovirus incidence rates. HAdV was detected at a higher rate in children in the preschool and school-age groups, and most cases were detected in the fall and winter seasons. The incidence rate of HAdV was negatively correlated with meteorological factors (temperature, hours of sunshine, and wind speed) and the atmospheric pollutant O₃, and positively correlated with atmospheric pollutants (CO, NO₂, and SO₂). The influence of these pollutants on the prevalence of HAdV infection should not be ignored.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-24515-5.

The ability to detect nucleic acids at ever-lower detection limits is crucial in molecular science, enabling disease management to shift from reactive response to proactive prediction and personalized medicine. This review synthesizes the technological landscape, which is organized into three strategies. First, we examine amplification-dependent platforms, charting their evolution from the quantitative polymerase chain reaction (PCR) to the absolute quantification of digital PCR (dPCR) and the rapid point-of-care utility of isothermal methods. Second, we explore amplification-free paradigms, where technologies such as clustered regularly interspaced short-palindromic repeat (CRISPR)-based diagnostics and nanopore sequencing achieve sensitivity through signal generation mechanisms that directly interrogate native target molecules. Finally, we detail the emergence of nanotechnology-based biosensors, which employ the physicochemical properties of nanomaterials to transduce molecular recognition events into measurable optical or electronic signals. The clinical and scientific translation of these platforms is very useful, enabling noninvasive cancer monitoring via liquid biopsy, real-time infectious disease surveillance, and the reconstruction of past ecosystems from trace environmental DNA. However, realizing the full potential of these technologies is contingent on overcoming significant translational hurdles. Critical challenges include the standardization of preanalytical workflows to ensure sample integrity and the engineering of fully integrated, cost-effective "sample-in, answer-out" systems suitable for widespread deployment. The future trajectory of the field points toward the convergence of these molecular tools with microfluidics, automation, and artificial intelligence, thus promoting decentralized and predictive healthcare.

Recent viral outbreaks have shown the need for reliable diagnostic platforms to rapidly detect various viral and bacterial pathogens at the point-of-care. Over the last decade, isothermal nucleic acid amplification methods have emerged as an appealing alternative to standardized polymerase chain reaction (PCR) tests due to their high sensitivity, selectivity, low cost, and simple assay setup. Virtually all nucleic acid testing platforms require a labor-intensive sample preparation step, limiting the scalability and usability of recent alternatives. This article describes multiplexed isothermal detection of respiratory viruses on a valve-free, autonomously loading microfluidic platform-VirChip. We demonstrate that an optimized loop-mediated isothermal amplification (LAMP) enables the simultaneous detection of SARS-CoV-2, influenza A, influenza B, and RSV (A/B) with a limit of detection of 100 RNA copies per reaction. The platform is highly selective, as no cross-reactivity amongst the targeted pathogens was observed in patient samples. Furthermore, crude nasal swab samples can be directly applied to the chip, eliminating the requirement for expensive and laborious RNA isolation and sample workup. VirChip facilitates rapid, inexpensive, and multiplexed detection, allowing pathogen screening by primary care providers not only in hospitals but also in resource-limited areas.

Shrimp shell flavoring is a food additive used to give food flavor or umami taste. This umami taste comes from amino acids such as glutamate which are naturally found in shrimp shells and other foods that are rich in protein. This research aims to identify the effect of drying methods on glutamic acid levels, protein content, and the quality of sensory characteristics of shrimp shell flavorings. This research used an experimental method with a Completely Randomized Design (CRD). There were four types of treatment which were carried out six times. Treatment consists of drying method: drying temperature: drying duration, F1 (microwave: 120ºC: 120 minutes), F2 (oven: 100ºC: 90 minutes), F3 (cabinet: 100ºC: 120 minutes), F4 (dehydrator: 70ºC: 720 minutes). The glutamic acid test uses the UV-Vis Spectrophotometry method, the protein test uses the Kjeldahl method and the sensory characteristics use hedonic quality with 25 semi-trained panelists. Analysis of data from glutamic acid used Kruskal Wallis continued with Man Whitney, protein tests used ANOVA, sensory characteristics used the Friedman Test and continued with the Wilxocon Test. Variations in drying methods had a significant effect on glutamic acid levels (p=0.002). The results of protein analysis showed no influence on variations in drying methods with a value of (p=0.131). The sensory characteristics of color (p=0.00), taste (p=0.002) while the parameters of texture (p=0.055), aroma (p=0.246) did not show any influence. The highest glutamic acid content was oven dried with a level of 226.5%/g. The highest protein content in dehydrator drying was 41.38%/g.

Beta-thalassemia is a single-gene recessive disorder caused by mutations in the HBB gene, and approximately 1.5% of the global population are carriers of β-thalassemia. It is therefore vital to establish a rapid and sensitive method to detect the mutant genes of β-thalassemia. In this study, a CRISPR/Cas12a-mediated amplification-free surface-enhanced Raman spectroscopy (SERS) biosensor was developed. This biosensor uses sea urchin-shaped gold nanoparticles (SUGNPs) as the SERS enhancement substrate and 4-mercaptobenzoic acid (4-MBA) as the Raman reporter. It couples the SUGNPs/4-MBA with magnetic beads through single-strand DNA (ssDNA) to form an SERS probe with magnetic responsiveness. The presence of the β-thalassemia target mutation gene CD31 activates the cleavage activity of Cas12a, leading to non-specific cleavage of single-stranded DNA (ssDNA) on the probe. This results in a significant reduction in SERS intensity. This signal change enables quantitative detection of the target gene, thereby significantly enhancing the sensitivity of nucleic acid testing. We employed magnetic separation technology to enrich target nucleic acids in serum while eliminating matrix interference, enabling specific recognition and quantitative detection of the mutated CD31 gene. This method exhibits excellent linearity over a concentration range 0.1 fM to 10pM, with a detection limit of 0.1 fM and a detection time of only 40min. Compared to traditional qPCR and other CRISPR methods, this approach is simple, rapid, and offers advantages such as high sensitivity, high specificity, and cost-effectiveness. By simply replacing the crRNA, it can detect multiple β-thalassemia and other disease genes, demonstrating broad clinical application potential.