Recombinase polymerase amplification (RPA) is a promising technology for amplifying nucleic acids at a constant temperature; its simple procedure, minimal equipment requirements, and fast detection times make it highly suitable for food safety applications. This study develops two rapid and sensitive assays for detecting Yersinia pseudotuberculosis in food based on RPA technology. The first assay combines RPA with a lateral flow stick (RPA-LFS) for visual readouts. The method showed high specificity against common food-borne bacteria and could detect Y. pseudotuberculosis at temperatures between 30 and 45 °C. High concentrations of templates were detected within 10 min, whereas low concentrations required 20-30 min. The limit of detection (LOD) with 95% probability by probit analysis was determined to be 206 CFU/mL. The second assay integrates RPA with real-time fluorescence detection (RT-RPA), allowing continuous monitoring of amplification products. RT-RPA detected as few as 47.5 copies/μL for pure DNA templates and 103 CFU/mL for bacteria suspensions. Amplification occurred in 3-14 min for 106-102 copies/μL of DNA template and 3-10 min for 107-103 CFU/mL of boiled bacterial suspensions. Both methods showed 100% accuracy in artificially contaminated pork samples and produced results consistent with qPCR. The RPA-LFS and RT-RPA assays constructed in this study can rapidly and accurately detect Y. pseudotuberculosis, which is crucial for food safety emergencies.

There have been no previous studies on male HPV infection in the Huizhou region. This research aims to investigate the HPV infection rate and genotype distribution among male patients in this area, offering valuable insights for developing targeted preventive strategies against HPV infection in male population. This study included 1009 male patients from Huizhou Central People’s Hospital who underwent HPV genotype testing between 2014 and 2023. We analyzed the distribution of HPV genotypes by year, age group, and diagnosis. Additionally, clinical data from 308 HPV-positive patients were retrospectively collected, and differences in high-risk vs. low-risk types, single vs. multiple infections, and genotype correlations were analyzed. The overall HPV positivity rate was 30.53

Whooping cough, caused by the highly contagious Bordetella pertussis (B. pertussis), poses a significant public health risk, particularly among unvaccinated children. Following the COVID-19 pandemic, an increase in B. pertussis cases has underscored the urgent need for rapid and reliable diagnostic tools to mitigate its transmission. This research introduces an innovative method for detecting B. pertussis that employs recombinase polymerase amplification in combination with a nanoparticle-based lateral flow biosensor (RPA-LFB). The assay can be completed in about 45 min, is easy to perform, and requires no specialized equipment. It demonstrated a detection limit of 10 copies per reaction and 100% analytical specificity, with no cross-reactivity to other common respiratory pathogens. Clinical assessment of 58 samples revealed a sensitivity of 85.7%, a specificity of 100%, and a positive predictive value of 100%. This field-deployable, instrument-free method is particularly advantageous for use in resource-limited or primary care settings, offering a practical solution for the timely diagnosis and control of B. pertussis infections.

BackgroundPersistent high-risk HPV infection is the major cause of cervical carcinogenesis, but the clinical relevance of Chlamydia trachomatis (CT) coinfection remains uncertain. We examined the epidemiological patterns of CT–HPV coinfection and their associations with cervical lesion severity among women in Southern China.MethodsThis retrospective study included 21,275 outpatients who underwent concurrent HPV genotyping, CT PCR testing, and cytological or histopathological evaluation between 2015 and 2024. Age-specific patterns and temporal trends were assessed. Age-adjusted logistic regression was used to examine associations of HPV and CT with cervical lesion categories. Six DNA methylation markers were analyzed in a subset of samples.ResultsOverall prevalence was 25.7% for HPV, 8.7% for CT, and 3.8% for HPV–CT coinfection, with declining trends over time. CT infection and HPV–CT coinfection were more common in younger women. HPV positivity was significantly higher in CT-positive than CT-negative women (43.9% vs 23.9%, p < 0.001). CT infection and coinfection were more frequently observed in ASC-US/-H and LSIL, while CT showed weaker associations with advanced lesions and was not significantly associated with invasive cervical carcinoma. Among the six methylation markers, only PAX1 showed significant differences among the three groups, while HPV–CT coinfection was not associated with additional methylation alterations compared with HPV infection alone.ConclusionsCT infection and HPV–CT coinfection were concentrated in younger women and were more frequently associated with low-grade cervical abnormalities. CT positivity was also associated with a higher prevalence of HPV infection.

Echovirus 18 (E18) has re-emerged as a global public health concern in recent years because of its association with severe neonatal systemic diseases that pose a risk of high mortality. The lack of effective intervention strategies for E18 infections is largely attributed to limited knowledge regarding molecular epidemiology and recombination patterns. In this study, we obtained seven full-length E18 sequences from infants in Guangdong Province and combined them with representative sequences from GenBank. Using this expanded dataset, we analysed the molecular epidemiological features, genetic characteristics, and recombination patterns of E18. Global statistics reveal a distinct double-peak pattern in the frequency of E18 infections throughout the year in the Northern Hemisphere. All strains isolated from clinical specimens were classified as genotype C2, which has emerged as the predominant circulating strain in Guangdong Province and globally. Several potential recombination events with E30 were identified among these seven strains, particularly in the P2 and P3 non-structural regions. This study offers new insights into the global dissemination, genetic diversity, and phylodynamics of E18, potentially providing valuable information for designing antiviral vaccines and the implementation of sustainable surveillance strategies to enhance virus prevention and control during public health crises.

Human adenovirus (HAdV) infections are prevalent and can lead to severe respiratory, ocular, and gastrointestinal diseases, thereby posing a significant public health risk, particularly among children and immunocompromised individuals. The genetic diversity of HAdV complicates rapid and accurate detection, highlighting the necessity for efficient and straightforward diagnostic methods. This research introduces a convenient and rapid one-pot RPA-CRISPR/Cas13a diagnostic system that is both sensitive and specific for the identification of HAdV DNA. The system incorporates two detection modalities: RPA-CRISPR/Cas13a-Flu for fluorescence-based detection and RPA-CRISPR/Cas13a-LFS for visual detection. The sensitivity of the RPA-CRISPR/Cas13a-Flu method was measured at 5 copies per reaction, while the RPA-CRISPR/Cas13a-LFS demonstrated a sensitivity of 50 copies per reaction, with both methods achieving 100 % specificity. Validation against 175 clinical samples indicated that the RPA-CRISPR/Cas13a-Flu had a sensitivity of 95.95 % (95 % CI 88.75-98.61), specificity of 100 % (96.34-100), accuracy of 98.29 % (95.08-99.42), and a Kappa coefficient of 0.965 when compared to traditional qPCR methods. The RPA-CRISPR/Cas13a-LFS exhibited 81.08 % (70.71-88.38) sensitivity, 100 % (96.34-100) specificity, 92.00 % (87.02-95.17) accuracy, and a Kappa coefficient of 0.832. Our developed onepot RPA-CRISPR/Cas13a system offers a rapid, user-friendly, and visually interpretable alternative to PCR for onsite HAdV detection, with potential applications in early diagnosis, subtyping, drug resistance detection, and portable monitoring of various infectious pathogens.

The epidemiological and clinical aspects of Human Papillomavirus (HPV) infection in women have been extensively studied. However, there is a lack of information regarding HPV characteristics in males. In this study, we conducted a retrospective and observational study of 3737 consecutive male individuals attending outpatient clinics of Guangdong Women and Children Hospital from 2012 to 2023 in Guangzhou, South China, to determine the age- and genotype-specific prevalence of HPV in men. The results showed the overall prevalence of HPV among men was 42.15% (1575/3737), with variations ranging from 29.55% to 81.31% across distinct diagnostic populations. Low-risk HPV6 (15.47%), HPV11 (8.94%), and high-risk HPV52 (5.51%) were the most common types. The annual HPV prevalence decreased significantly (Z = -3.882, p < .001), ranging from 31.44% to 52.90%. 28.77% (1075/3737) of men manifested infection with a singular HPV type, predominantly identified as a low-risk type. The age-specific distribution of HPV infections revealed distinctive peaks in the < 25 y age group (47.60%, 208/437) and the 40-44 y age group (44.51%, 154/346). Notably, the positive rate of Chlamydia trachomatis was significantly higher among HPV-positive individuals in comparison to HPV-negatives (16.14% vs. 11.25%, p < .05). Our findings reveal a substantial prevalence of HPV infection among outpatient men in Guangzhou, South China. It is recommended to consider the inclusion of HPV vaccination for adolescent males in national immunization schedules, once an adequate supply of vaccines is accessible.

Background: Enteroviruses (EV) are common and can cause severe diseases, particularly in young children. However, the information of EV infection in infants in China is limited due to the vast population size and extensive geographical area of the country. Here, we conducted a retrospective multicenter analysis of available EV data to assess the current epidemiological situation in the infant population in southern China. Methods: The study enrolled infants with suspected EV infection from 34 hospitals across 12 cities in southern China between 2019 to 2022, and the confirmation of EV was done using RT-PCR and VP1 gene sequencing. Results: Out of 1221 infants enrolled, 330 (27.03%) were confirmed as EV-infected. Of these, 260 (78.79%) were newborns aged 0-28 days. The EV belonged to three species: EV-B (80.61%), EV-A (11.82%), and human rhinovirus (7.58%). Newborns were more susceptible to EV-B than older infants (p < 0.001). Within EV-B, we identified 15 types, with coxsackievirus (CV) B3 (20.91%), echovirus (E) 11 (19.70%), and E18 (16.97%) being the most common. The predominant EV types changed across different years. EV infection in infants followed a seasonal pattern, with a higher incidence from May to August. Furthermore, perinatal mother-to-child EV transmission in 12 mother-newborn pairs were observed. Conclusion: Our study is the first to demonstrate the emergence and widespread circulation of EV-B species, mainly CVB3, E11, and E18, in southern China, primarily affecting young infants. This research provides valuable insights for future epidemic assessment, prediction, as well as the elimination of mother-to-child transmission.

Sexually transmitted diseases (STDs) are usually caused by co-infections of bacteria and viruses. However, there is a lack of products that possess both antibacterial and antiviral activities without using chemical drugs. Here, we developed a carrageenan silver nanoparticle composite hydrogel (IC-AgNPs-Gel) based on the antiviral activity of iota carrageenan (IC) and the antibacterial effect of silver nanoparticles (AgNPs) to prevent STDs. IC-AgNPs-Gel showed excellent biocompatibility, hemostasis, antibacterial and antiviral effects. IC-AgNPs-Gel not only effectively prevented S. aureus, E. coli, P. aeruginosa, and C. albicans without using antibiotics, but also significantly inhibited human papilloma virus (HPV)-16 and HPV-6 without using chemotherapy drugs. Moreover, IC-AgNPs-Gel showed the effects of accelerating infected wound healing and reducing inflammation in a rat wound model infected with S. aureus. Therefore, the multifunctional hydrogel shows great potential application prospect in preventing STDs.

Persistent high-risk human papillomavirus (HPV) infection is the pivotal cause of cervical carcinogenesis. HPV types distribution varies greatly by region, and its long-term changes of prevalence remain to be fully characterized in China. Here, the largest population of 198,111 consecutive women who underwent routine cervical screening were investigated from 2015 to 2021 in Guangzhou, south China. The results showed that the overall HPV prevalence was 21.66% (42,911/198,111), and the annual prevalence increased significantly from 2015 to 2021 (p < 0.001). HPV52, 16, 58, CP8304, 51, 53, 39, and 68 were the most prevalent HPV types. The relative HPV-positive rate correlated positively with the progression of cervical intraepithelial neoplasia (p < 0.001); HPV16 was the predominant carcinogenic type, followed by HPV52 and HPV18. HPV infections were significantly age-specific, and 26.51% (11,375/42,911) of cases were caused by multiple HPV types. In addition, HPV infections typically cleared over a median time of 16 (interquartile range 9-31) months, and the clearance of HPV16 was significantly faster than that of other types (p < 0.001). These findings may serve as a guide for local governments to evaluate HPV vaccination and cervical cancer prevention strategies in south China.

Abstract Background Enterovirus (EV) infections are being increasingly seen in younger infants, often being more severe than in older children. The risk factors of EV infection in infants have been inadequately investigated till date. Methods We conducted a retrospective study on hospitalized children with laboratory-confirmed EV infection (50 infants aged 0–3 months and 65 older than 3 months) at a tertiary care center in China. Prevalence, clinical characteristics, and genetic features of the virus were analyzed, and independent predictors for severe infection were assessed. Results Clinical findings showed that severe infection was more common in infants aged 0–3 months than in older children (78.0% vs. 35.4%, p < 0.001), with higher morbidity of pneumonia, meningitis, and sepsis (p < 0.01). EV-B types were detected more frequently in infants aged 0–3 months than in older children (88.0% vs. 7.7%, p < 0.001). Echovirus 11 was the most identified EV-B, and it recombined with E6 in P2 and P3 regions. Risk factors for severe EV infection included EV-B types infection, age less than 3 months, elevated alanine aminotransferase level, abnormal platelet count, and abnormal cerebrospinal fluid characteristics. Conclusions Our data indicated that EV-B types mainly cause severe infection in infants aged 0–3 months. Therefore, knowledge about EV-B types could have implications in designing effective intervention and prevention strategies for young infants with severe EV infection.

Background: Genital Chlamydia trachomatis (CT) is one of the most common agents of sexually transmitted infections and can cause severe disorders. This study aimed to analyse the genetic and clinical characteristics of genital CT infection among women in Guangzhou, China. Methods: From September 2020 to August 2021, a total of 8955 female patients were enrolled in this study. The presence of genital CT was detected by real-time PCR, and 273 positive samples were randomly selected for further genetic and clinical characteristics analysis. Results: The positive rate of genital CT infection was 7.5% (670/8955), with the highest rate in women aged 21-30 years. A total of 8 genotypes were identified: D-H, J, K, and recombinant genotype Ba/D. The predominant genotype was J (n = 78, 28.6%), followed by E (n = 63, 23.1%), F (n = 48, 17.6%), and D (n = 38, 13.9%). Abnormal vaginal discharge (n = 165, 61.8%), cervical columnar epithelial ectopy (n = 124, 46.4%), vaginal itching (n = 77, 28.8%), and lower abdominal pain (n = 61, 22.8%) were the predominant symptoms. Additionally, genotype G infection exhibited a significantly higher rate of abnormal vaginal discharge (P = 0.03) and genotype D infection exhibited a higher white blood cell count (P = 0.01) than the other genotypes. Phylogenetic analysis revealed a total of 20 variants with 25 mutation positions and the H2 variant in four patients was first discovered in our study. Conclusions: Genotypes J, E, F, and D were the major genotypes of genital CT in Guangzhou, and they manifested as abnormal vaginal discharge, cervical columnar epithelial ectopy, vaginal itching, and lower abdominal pain. The present study provides guidance for future integrated interventions to reduce the burden of genital CT infection and accelerate the development of vaccines.

Genogroup II genotype 4 (GII.4) norovirus causes acute gastroenteritis in children, and its infection is more severe than that of other genotypes. Early and precise detection and treatment are critical for controlling its spread and reducing the severity of infection. In this study, a rapid and efficient isothermal assay for the GII.4 norovirus detection (GII.4-CRISPR detection) was developed based on the CRISPR/Cas13a system. The assay can be applied without expensive instrumentation, and the results can be read via both fluorescence and lateral flow strip (LFS). The analytical sensitivity of this assay was 5 copies/reaction, and there was no cross-reaction with other genotypes of norovirus or other clinically common pathogens. There was a coincidence rate of 100% between our assay and commercial quantitative polymerase chain reaction. GII.4-CRISPR detection improves upon the shortcomings of some previously established molecular methods of detection, particularly with regard to accessibility. It provides an alternative tool for outbreak control and early diagnosis of GII.4 norovirus infection.

The white spot syndrome virus is the most destructive virus threatening the shrimp industry worldwide, causing hundreds of millions of dollars in economic losses each year. There is currently no specific medicine to treat it. Therefore, rapid and accurate detection of WSSV is of great significance for controlling its spread and reducing economic losses. Traditional detection methods, such as polymerase chain reaction (PCR) and quantitative fluorescent PCR, rely on laboratory equipment and are not suitable for field testing. In this study, recombinase polymerase amplification (RPA) combined with a lateral flow strip (LFS) was developed. This method targets the entire genome and designs primers and probes accordingly. The detection can be completed in 30 min at 37°C, and the detection limit of each reaction is 20 copies, which is much more sensitive than other detection methods. The RPA-LFS method is highly specific to the white spot syndrome virus and has no cross-reactivity with other common shrimp viruses or pathogens. In total, 100 field samples were tested and compared to the real-time PCR method. Both methods detected 8 positive results, and the positive detection rate was 100%. The method was fast, simple, specific, and sensitive. It does not rely on laboratory equipment and has broad application prospects for in-field detection, especially in remote areas with underdeveloped medical equipment.

Acute hepatopancreatic necrosis disease (AHPND) is a significant deadly infectious disease in the shrimp farming industry, causing serious economic losses globally every year. Because of the rapid progress speed, lack of effective treatment and high mortality rate of AHPND, monitoring with frequent diagnostic tests is vital for a successful prevention. The conventional histopathological diagnosis fell far short of the requirement for efficient monitoring, and the polymerase chain reaction (PCR)-based molecular diagnostic methods that rely on sophisticated thermocycler and trained personnel are hardly applicable in the field. Combining the recombinase polymerase amplification (RPA) and the lateral flow strips (LFSs), a diagnostic method suitable for on-site everyday monitoring of AHPND has been established in this study. This RPA-LFS method targeted the binary toxic photorhabdus insect-related genes PirA and PirB on a virulence plasmid of the AHPND-causative Vibrio parahaemolyticus strains. The diagnostic test was completed within 30 min at 37°C and showed good specificity and good sensitivity of 20 fg DNA of the AHPND shrimp or one colony-forming unit of the causative bacterium per reaction, which was better than the administration-approved standard AP4 assay. Crude templates from sample boiling could be directly used. Tests of clinical samples showed 100% consistency of this method with the standard AP4 assay. This RPA-LFS method can be a good choice for on-site diagnosis of AHPND with quick response time, easy procedure and low demand for resources, and should have significant value for the control of spreading of this dangerous disease in farmed shrimp.

Hand, foot, and mouth disease (HFMD) is a common infectious disease affecting mainly children under 5 years of age. Coxsackievirus A6 (CVA-6), a major causative pathogen of HFMD, has caused outbreaks in recent years. Currently, no effective vaccine or antiviral treatments are available. In this study, one-step reverse-transcription recombinase polymerase amplification (RT-RPA), combined with a disposable lateral flow strip (LFS) assay, was developed to detect CVA-6. This assay can be performed in less than 35 min at 37°C without expensive instruments, and the result can be observed directly with the naked eye. The sensitivity of the RT-RPA-LFS was 10 copies per reaction, which was comparable to that of the conventional real-time quantitative polymerase chain reaction (qPCR) assays. Moreover, the assay specificity was 100%. The clinical performance of the RT-RPA-LFS assay was evaluated using 142 clinical samples, and the coincidence rate between RT-RPA-LFS and qPCR was 100%. Therefore, our RT-RPA-LFS assay provides a simple and rapid approach for point-of-care CVA-6 diagnosis.

Acute hepatopancreatic necrosis disease (AHPND) is an emerging disease in the shrimp farming industry with a high mortality rate that causes serious economic losses in Asia, Africa, and America [1]. AHPND is also known as early mortality syndrome because of its early onset of the disease at 5–30 days of shrimp postlarvae stocking and a high mortality rate up to 100%. Symptoms of AHPND include lighter hepatopancreas, atrophy, jejunal empty stomach, softened carapace, and loss of vitality. AHPND is caused by infection of special strains of Vibrio parahaemolyticus (referred to VPAHPND in this study) containing the 69-kb pVA1 plasmid with the binary toxic Photorhabdus insect-related genes PirA and PirB (referred to PirAB in this study) [2]. In January 2016, the World Organization for Animal Health (OIE) added AHPND to the OIE List of Aquatic Animal Diseases. Rapid, sensitive, and reliable diagnosis assays for AHPND are...

BACKGROUND:Andrographolide (Andro), a diterpenoid extracted from Andrographis paniculata, has been shown to attenuate pulmonary fibrosis in rodents; however, the potential mechanisms remain largely unclear. This study investigated whether and how Andro alleviates bleomycin (BLM)-induced NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activation and epithelial-mesenchymal transition (EMT) in the lung epithelial cells.METHODS:The in vivo effects of Andro were evaluated in a rat model of BLM-induced pulmonary fibrosis. The roles of Andro in BLM-induced NLRP3 inflammasome activation, EMT and AKT/mTOR signaling were investigated using human alveolar epithelial A549 cells.RESULTS:We found that Andro significantly alleviated pulmonary edema and histopathological changes, decreased weight loss, and reduced collagen deposition. Andro downregulated the levels of NLRP3, the adaptor molecule apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 in the lungs of BLM-treated rats, suggesting the inhibitory effect of Andro on NLRP3 inflammasome activation in vivo. Additionally, the symptoms of BLM-mediated EMT phenotype in the lung were also attenuated after Andro administration. In vitro, Andro also markedly inhibited BLM-induced NLRP3 inflammasome activation and EMT in A549 cells. Moreover, Andro inhibited BLM-induced phosphorylation of AKT and mTOR in A549 cells, suggesting that AKT/mTOR inactivation mediates Andro-induced effects on BLM-induced NLRP3 inflammasome activation and EMT.CONCLUSIONS:These data indicate that Andro can reduce BLM-induced pulmonary fibrosis through suppressing NLRP3 inflammasome activation and EMT in lung epithelial cells via AKT/mTOR signaling pathway.

为建立副溶血弧菌快速检测方法,本研究以副溶血弧菌不耐热溶血素(tlh)基因作为靶序列设计特异性引物,经反应条件优化建立了检测副溶血弧菌的重组酶聚合酶扩增(RPA)方法.优化试验结果显示该方法在37℃和35 min条件下检测效果最佳;特异性试验结果显示,该方法除对副溶血弧菌的检测结果为阳性外,对其余8种常见的弧菌检测结果均为阴性,特异性较强;敏感性试验结果显示,该RPA方法对副溶血弧菌的检出限为104 cfu/mL,敏感性较高.利用建立的RPA检测方法、培养法及普通PCR方法同时对采集的48份海鲜样品进行检测,结果显示:该RPA方法能够对临床样品快速检测,并且检测结果与国家标准培养法及普通PCR方法的符合率为100％,能够满足临床样品的检测需求.本研究建立了一种检测副溶血弧菌的方法,该方法具有反应快速、特异性强、敏感性高等特点,为副溶血弧菌的快速检测奠定基础.