An improvement of real-time polymerase chain reaction system based on probe modification is required for accurate detection of African swine fever virus in clinical samples in Vietnam

Tran Ha Thi Thanh,Dang Anh Kieu,Ly Duc Viet,Vu Hao Thi,Hoang Tuan Van,Nguyen Chinh Thi,Chu Nhu Thi,Nguyen Vinh The,Nguyen Huyen Thi,Truong Anh Duc,Pham Ngoc Thi,Dang Hoang Vu 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.10

Objective: The rapid and reliable detection of the African swine fever virus (ASFV) plays an important role in emergency control and preventive measures of ASF. Some methods have been recommended by FAO/OIE to detect ASFV in clinical samples, including real-time polymerase chain reaction (PCR). However, mismatches in primer and probe binding regions may cause a false-negative result. Here, a slight modification in probe sequence has been conducted to improve the qualification of real-time PCR based on World Organization for Animal Health (OIE) protocol for accurate detection of ASFV in field samples in Vietnam. Methods: Seven positive confirmed samples (four samples have no mismatch, and three samples contained one mutation in probe binding sites) were used to establish novel real-time PCR with slightly modified probe (Y = C or T) in comparison with original probe recommended by OIE. Results: Both real-time PCRs using the OIE-recommended probe and novel modified probe can detect ASFV in clinical samples without mismatch in probe binding site. A high correlation of cycle quantification (Cq) values was observed in which Cq values obtained from both probes arranged from 22 to 25, suggesting that modified probe sequence does not impede the qualification of real-time PCR to detect ASFV in clinical samples. However, the samples with one mutation in probe binding sites were ASFV negative with OIE recommended probe but positive with our modified probe (Cq value ranked between 33.12-35.78). Conclusion: We demonstrated for the first time that a mismatch in probe binding regions caused a false negative result by OIE recommended real-time PCR, and a slightly modified probe is required to enhance the sensitivity and obtain an ASF accurate diagnosis in field samples in Vietnam.

The potential efficacy of the E2-subunit vaccine to protect pigs against different genotypes of classical swine fever virus circulating in Vietnam

Ha Thi Thanh Tran,Duc Anh Truong,Viet Duc Ly,Hao Thi Vu,Tuan Van Hoang,Chinh Thi Nguyen,Nhu Thi Chu,Vinh The Nguyen,Duyen Thuy Nguyen,Kohtaroh Miyazawa,Takehiro Kokuho,Hoang Vu Dang 대한백신학회 2020 Clinical and Experimental Vaccine Research Vol.9 No.1

Purpose: To date, many kinds of classical swine fever (CSF) vaccines have been developed to protect against this disease. However, the efficacy of these vaccines to protect the pig against field CSF strains needs to be considered, based on circulating strains of classical swine fever virus (CSFV). Materials and Methods: Recombinant E2-CSFV protein produced by baculovirus/insect cell system was analyzed by western blots and immunoperoxidase monolayer assay. The effect of CSFV-E2 subunit vaccines was evaluated in experimental pigs with three genotypes of CSFV challenge. Anti-E2 specific and neutralizing antibodies in experimental pigs were analyzed by blocking enzyme-linked immunosorbent assay and neutralization peroxidize-linked assay. Results: The data showed that CSFV VN91-E2 subunit vaccine provided clinical protection in pigs against three different genotypes of CSFV without noticeable clinical signs, symptoms, and mortality. In addition, no CSFV was isolated from the spleen of the vaccinated pigs. However, the unvaccinated pigs exhibited high clinical scores and the successful virus isolation from spleen. These results showed that the E2-specific and neutralizing antibodies induced by VN91-E2 antigen appeared at day 24 after first boost and a significant increase was observed at day 28 (p<0.01). This response reached a peak at day 35 and continued until day 63 when compared to controls. Importantly, VN91-E2 induced E2-specific and neutralizing antibodies protected experimental pigs against high virulence of CSFVs circulating in Vietnam, including genotype 1.1, 2.1, and 2.2. Conclusion: These findings also suggested that CSFV VN91-E2 subunit vaccine could be a promising vaccine candidate for the control and prevention of CSFV in Vietnam.

Efficacy of electrochemically activated water solution in gingivitis treatment

Dinh‑Toi Chu,Van‑Huy Pham,Thien Chu‑Dinh,Truong Nhu Ngoc Vo,Duc Long Duong,Van Nhon Bui,Minh Son Tong,Thi Thu Phuong Nguyen,Quynh Anh Le,Khanh‑Hoang Nguyen 한국약제학회 2019 Journal of Pharmaceutical Investigation Vol.49 No.3

Mouthwash is one of the most commonly oral healthcare products for prevention and treatment of gum diseases. Dr. ECA is a disinfectant solution manufactured by a Vietnamese company and has been licensed for oral healthcare, but its effect on gingivitis is still unclear. To evaluate the effectiveness of this product in the treatment of gingivitis, we conducted a clinical study in 60 students diagnosed with gingivitis to compare the results of gingivitis treatment using Dr. ECA and a control mouthwash (a demonstrated anti-gingivitis product). The data showed that in the treated group, the rate of good Gingivitis Index (GI) before treatment was 46.67%, but after 2 and 4 weeks of using Dr. ECA that rate was increased to 96.67% and 100%, respectively. The percentage of patients with bleeding on probing was 63.33% but declined sharply after 2 weeks of treatment to 6.67%, and further reduced to 3.33% after 4 weeks of treatment. The treatment outcomes in were similar in the Dr. ECA treated group and control one with no statistically significant difference. Our results suggest that the electrolyte saline solution Dr. ECA is effective in treating gingivitis. Its effects are similar to other mouthwash products that had been approved and marketed.

Genome‑wide identification, organization, and expression profiles of the chicken fibroblast growth factor genes in public databases and Vietnamese indigenous Ri chickens against highly pathogenic avian influenza H5N1 virus infection

Truong Anh Duc,Tran Ha Thi Thanh,Chu Nhu Thi,Nguyen Huyen Thi,부 티 하오,Hong Yeojin,송기덕,Dang Hoang Vu,홍영호 아세아·태평양축산학회 2023 Animal Bioscience Vol.36 No.4

Objective: Fibroblast growth factors (FGFs) play critical roles in embryo development, and immune responses to infectious diseases. In this study, to investigate the roles of FGFs, we performed genome-wide identification, expression, and functional analyses of FGF family members in chickens. Methods: Chicken FGFs genes were identified and analyzed by using bioinformatics approach. Expression profiles and Hierarchical cluster analysis of the FGFs genes in different chicken tissues were obtained from the genome-wide RNA-seq. Results: A total of 20 FGF genes were identified in the chicken genome, which were classified into seven distinct groups (A-F) in the phylogenetic tree. Gene structure analysis revealed that members of the same clade had the same or similar exon-intron structure. Chromosome mapping suggested that FGF genes were widely dispersed across the chicken genome and were located on chromosomes 1, 4-6, 9-10, 13, 15, 28, and Z. In addition, the interactions among FGF proteins and between FGFs and mitogen‑activated protein kinase (MAPK) proteins are limited, indicating that the remaining functions of FGF proteins should be further investigated in chickens. Kyoto encyclopedia of genes and genomes pathway analysis showed that FGF gene interacts with MAPK genes and are involved in stimulating signaling pathway and regulating immune responses. Furthermore, this study identified 15 differentially expressed genes (DEG) in 21 different growth stages during early chicken embryo development. RNA-sequencing data identified the DEG of FGFs on 1- and 3-days post infection in two indigenous Ri chicken lines infected with the highly pathogenic avian influenza virus H5N1 (HPAIV). Finally, all the genes examined through quantitative real-time polymerase chain reaction and RNA-Seq analyses showed similar responses to HPAIV infection in indigenous Ri chicken lines (R2 = 0.92– 0.95, p<0.01). Conclusion: This study provides significant insights into the potential functions of FGFs in chickens, including the regulation of MAPK signaling pathways and the immune response of chickens to HPAIV infections.