EvaGreen-based real-time PCR assay for sensitive detection of enzootic nasal tumor virus 2

Huang, Qingyuan; Ye, Chao; Chen, Tingting; Jiang, Jiali; Peng, Yuanyi; Chen, Jing; Fang, Rendong

doi:10.1016/j.mcp.2019.02.003

Cited by 5 publications

(4 citation statements)

References 18 publications

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“…Recent studies indicated that the incidence of ENA in China showed an upward trend, and its spreading areas were also expanding. In recent years, ENA has been reported in multiple provinces in China, including Shaanxi, Anhui, Sichuan, Chongqing, Fujian, Guangxi, Guangdong, and Inner Mongolia [ 23 ]. Hence, the genetic variation and evolution of ENTV-2 in China need to be further investigated.…”

Section: Discussionmentioning

confidence: 99%

Genomic Sequencing and Analysis of Enzootic Nasal Tumor Virus Type 2 Provides Evidence for Recombination within the Prevalent Chinese Strains

Li,

Niu,

Liu

et al. 2024

Veterinary Sciences

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In recent years, the clinical cases of ENTV-2 infection have increased and become prevalent in several provinces of China. In this study, we reported the occurrence of ENTV-2 in one goat farm in Chongqing, southwest China. The complete genome of an emerged ENTV-2 isolate (designated as CQ2) was sequenced with 7468 bp in length. Phylogenetic analysis revealed that ENTV-2 consisted of two main lineages. Lineage 1 was composed of Chinese strains and could be subdivided into five sublineages. CQ2 and the other six recent isolates from China were clustered in sublineage 1.5; however, CQ2 was significantly different from the other six isolates. Furthermore, recombination analysis suggested that CQ2 might be a recombinant variant derived from sublineage 1.5 and sublineage 1.2 strains, with the recombination region in areas of pro and pol genes. In conclusion, we sequenced and analyzed the complete genome of a potential ENTV-2 recombinant, which may contribute to our understanding of the genetic variation and evolution of ENTV-2 in China.

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Section: Discussionmentioning

confidence: 99%

Genomic Sequencing and Analysis of Enzootic Nasal Tumor Virus Type 2 Provides Evidence for Recombination within the Prevalent Chinese Strains

Li,

Niu,

Liu

et al. 2024

Veterinary Sciences

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“…In comparison with SYBR Green, EvaGreen has some advantages. Firstly, it is more sensitive, and secondly, due to its less negative influence on reaction, the occurrence of non-specific amplification is rare (18)(19)(20).…”

Section: Discussionmentioning

confidence: 99%

Designing and Development of Simultaneous Detection of Neisseria meningitidis and Streptococcus pneumoniae based on EvaGreen Real-Time PCR

Shokrpour

Alebouyeh

Fallah³

et al. 2022

Arch Clin Infect Dis

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Background: Neisseria meningitidis and Streptococcus pneumoniae are serious causes of invasive infections associated with high mortality and morbidity worldwide, particularly meningitis. Efficient diagnostic strategies play a crucial role in the management of disease and the prevention of overtreatment. The low sensitivity and time-consuming nature of culture and gram stain methods have led to the demand for alternative methods in clinical laboratories. Objectives: This study aims to design and develop a rapid, sensitive, and cost-effective EvaGreen-based real-time PCR to simultaneously detect N. meningitidis and S. pneumoniae. Methods: We designed and evaluated an accurate, reliable, and inexpensive approach based on EvaGreen dye real-time PCR to simultaneously detect N. meningitidis and S. pneumoniae in a single tube from cerebrospinal fluid. In a single-tube reaction, melting curve analysis was used to differentiate the amplicons of each pathogen. Analytical sensitivity and specificity of the assay were conducted by reference bacterial strains genomes. Besides, in order to clinical validation we used 53 positive CSF samples and 7 negative CSF samples. Results: Our assay demonstrated no amplification curve with non-target microorganisms indicating 100% analytical specificity. In the EvaGreen multiplex assay, the lower limit of detection (LLD) was nine copies/reaction for N. meningitidis and 13 copies/reaction for S. pneumoniae. The clinical validation of positive CSF samples revealed 100% sensitivity and no false positives. The reproducibility and repeatability of tested replicates indicated low intra-assay and inter-assay CVs of less than 1.5%. Conclusions: EvaGreen-based multiplex real-time PCR offers a rapid, affordable, and appropriate diagnostic tool to identify the main cause of bacterial meningitis.

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“…Currently, there are no commercially available diagnostic tools providing a non-invasive antemortem diagnosis of the disease; a RT-PCR on RNA extracted from nasal fluids has recently been developed (Huang et al, 2019). Due to the lack of circulating antibodies, serological screening for the virus seems impossible and thus the prevalence and genotypic distribution of ENTV are still unknown (Huang et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

First case of nasal transitional carcinoma in a goat infected with Enzootic Nasal Tumor Virus in Belgium

Delaude¹,

Raes²,

Leroux³

et al. 2020

VDT

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An eleven-year-old, female goat was presented for evaluation of breathing difficulties and epistaxis. Radiographs and computed tomographic (CT) examination of the head revealed the presence of a space-occupying lesion involving the right nasal passage and invading the cranial vault and retrobulbar space. Histologic examination and detection of viral genome from the nasal mass led to the diagnosis of nasal transitional carcinoma with concomitant infection with Enzootic Nasal Tumor Virus (ENTV-2). In this case report, the presence of a nasal transitional carcinoma is described in a goat; a very rare tumor which, to the authors’ knowledge, has not been previously reported in goats. Reverse transcription polymerase chain reaction (RT-PCR) detected the genome of ENTV-2 within the neoplastic tissue, suggesting an infectious etiology.

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EvaGreen-based real-time PCR assay for sensitive detection of enzootic nasal tumor virus 2

Cited by 5 publications

References 18 publications

Genomic Sequencing and Analysis of Enzootic Nasal Tumor Virus Type 2 Provides Evidence for Recombination within the Prevalent Chinese Strains

Genomic Sequencing and Analysis of Enzootic Nasal Tumor Virus Type 2 Provides Evidence for Recombination within the Prevalent Chinese Strains

Designing and Development of Simultaneous Detection of Neisseria meningitidis and Streptococcus pneumoniae based on EvaGreen Real-Time PCR

First case of nasal transitional carcinoma in a goat infected with Enzootic Nasal Tumor Virus in Belgium

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