The microRNA MiR-24-3p suppresses cancer progression by targeting TRIM11. The long noncoding RNA LUADT1 has been reported to promote lung adenocarcinoma proliferation. We found LUADT1 may form base pairing with miR-24-3p. This study aimed to explore the interactions among LUADT1, miR-24-3p, and TRIM11 in mantle cell lymphoma (MCL). Our study recruited 40 MCL patients and 40 healthy volunteers. Tumor tissues were collected from 40 newly diagnosed MCL patients and embedded in paraffin wax. B lymphocytes were isolated from all tissue samples by using CD19⁺ magnetic beads and DETACHaBEAD CD19. Human MCL cell line Grante-519 and JeKo-1 were transfected with LUADT1 and TRIM11 expression vectors, microRNA mimics or inhibitors. Then, quantitative polymerase chain reaction and Western blot were used to detect the level of relative messenger RNA and protein expression, respectively. Flow cytometry was performed to detect the apoptosis rate. LUADT1 and miR-24-3p were upregulated while TRIM11 was downregulated in MCL both in tissues and cell lines compared with hyperplastic lymphadenitis and peripheral lymphocyte cells. Bioinformatics analysis showed that LUADT1 may bind miR-24-3p, which can target TRIM11. Correlation analysis showed that LUADT1 was not significantly correlated with miR-24-3p. However, it was positively and significantly correlated with TRIM11. In MCL cells, LUADT1 overexpression led to upregulated TRIM11, whereas miR-24-3p overexpression led to downregulated TRIM11. Cell apoptosis analysis showed that LU-ADT1, miR-24-3p inhibitor and TRIM11 overexpression led to decreased apoptotic rate of MCL cells, whereas miR-24-3p overexpression led to an increased apoptotic rate of MCL cells. In addition, miR-24-3p overexpression attenuated the effects of LUADT1 overexpression. Therefore, LUADT1 was upregulated in MCL and could modulate TRIM11 by sponging miR-24-3p to inhibit cancer cell apoptosis.

To establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the M gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) reaction mixture was developed. A simple and rapid RT-qPCR detection method for PRRSV was developed by optimizing nucleic acid amplification conditions. The results showed that the method was able to specifically detect PRRSV without cross-reactivity with the other 11 porcine susceptible viruses. The sensitivities of the assay were 3.12 × 10⁰ copies/μL and 10⁰ TCID₅₀/μL for M gene and virus, respectively, and the repeatability and reproducibility (relative standard deviation, CV) of the assay were less than 2.5%. Based on the new fullly pre-mixed RT-qPCR reaction mixture, the RT-qPCR detection method may provide a new, simple, and rapid method for accurately detecting PRRSV.

The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 10⁹–6.43 × 10¹ copies/µL, with an R² value of 0.996 and a standard curve equation of y = −3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings.

As an infectious disease that poses a significant threat to the rapidly growing pig breeding industry, the detection of Haemophilus parasuis (HPS) is often compromised by various interfering substances present in the test sample during quantitative real-time PCR (qPCR). The rapid detection of HPS is important for the isolation of infectious pigs and their treatment. We designed and optimized a rapid qPCR test to detect the INFB gene of HPS in clinical and environmental samples on pig farms. The method was evaluated for its specificity, sensitivity, repeatability, anti-interference capability, and its ability to detect HPS in clinical samples. The results indicated that the method was specific for the detection of HPS when evaluated against pathogens and intestinal probiotics found in pig farms. By using a seven-fold dilution series of the recombinant plasmid DNA in triplicate, it was determined that the lowest limit of detection (LOD) for this method was less than 10 copies/µL. The results of inter-batch and intra-batch repeatability tests showed that the coefficient of variation (CV) was consistently below 1%. Furthermore, the impact of 14 endogenous and exogenous interfering substances on the Ct values detected by the HPS qPCR was found to be less than 5% when compared to the Ct values obtained in the absence of interfering substances. A total of 248 clinical samples were analyzed using the HPS qPCR, commercial kits, and corresponding national standards, yielding positive rates of 9.27%, 6.05%, and 9.27%, respectively. Notably, the positive and negative percent agreement between the detection method developed in this research and the national standard was 100%. These findings demonstrate that the established detection method is suitable for epidemiological research on HPS and for diagnosing clinical samples containing interfering substances, thereby providing essential technical support for the prevention and control of HPS.

To establish a rapid real-time RT-PCR method for differentiating wild-type classical swine fever virus (CSFV) strains from vaccine strains (HCLV), we designed a universal primer targeting the NS3 gene to detect wild-type CSFV strains and vaccine strains simultaneously, and two TaqMan-MGB probes were designed to differentiate between wild-type and vaccine strains. After optimizing the RT-qPCR conditions, a rapid dual TaqMan-MGB RT-qPCR method for the detection and identification of CSFV and HCLV was developed. The results showed that method could specifically detect CSFV and HCLV with no cross-reactivity with other swine pathogens. The analytic sensitivity for the NS3 gene of CSFV and HCLV were 1.67 × 10¹ copies/μL, respectively. For precision testing, the repeatability and reproducibility of the test was less than 2%. This method was successfully used for the rapid detection of 193 biological samples collected from CSFV-vaccinated pigs. This fast and accurate detection technology can be used for the detection of CSFV and is suitable for differentiating between wild-type CSFV strains and vaccine strains.