Development of a real‐time PCR assay for detection of African swine fever virus with an endogenous internal control

2020 ◽  
Vol 67 (6) ◽  
pp. 2446-2454 ◽  
Author(s):  
Yin Wang ◽  
Lizhe Xu ◽  
Lance Noll ◽  
Colin Stoy ◽  
Elizabeth Porter ◽  
...  
Keyword(s):  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Pcr Assay

Development of a triplex real-time PCR assay for detection and differentiation of gene-deleted and wild-type African swine fever virus

2020 ◽  
Vol 280 ◽  
pp. 113875
Author(s):  
Yanxing Lin ◽  
Chenfu Cao ◽  
Weijun Shi ◽  
Chaohua Huang ◽  
Shaoling Zeng ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Wild Type ◽  
Pcr Assay

scholarly journals Development of Real-Time PCR Based on A137R Gene for the Detection of African Swine Fever Virus

2021 ◽  
Vol 8 ◽  
Author(s):  
Dan Yin ◽  
Renhao Geng ◽  
Hui Lv ◽  
Chunhui Bao ◽  
Hongxia Shao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Economic Losses ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Taqman Pcr ◽  
Control And Eradication

African swine fever virus (ASFV) can infect domestic pigs and wild boars and causes huge economic losses in global swine industry. Therefore, early diagnosis of ASFV is important for the control and eradication of African swine fever (ASF). In this study, a SYBR Green-based real-time polymerase chain reaction (PCR) assay targeting the viral encoded A137R gene was established for the detection of ASFV infection. For the evaluation of the established real-time PCR, 34 clinical samples were assessed by both the A137R gene-based real-time PCR and OIE-recommended TaqMan PCR. The results showed that 85.29% (29/34) were detected by A137R gene-based real-time PCR, but only 79.41% (27/34) positive using OIE-recommended TaqMan PCR. Moreover, no cross-reaction with other common swine pathogens was found in the A137R gene-based real-time PCR. These results demonstrated that the established real-time PCR assay in this study showed better performance than the OIE-recommended method in detecting ASFV from clinical samples, which could be applied for control and eradication programs of ASF.

scholarly journals Development of a novel real-time PCR assay for rapid detection of African swine fever virus (ASFV) strains circulating in Vietnam

2020 ◽  
Author(s):  
Thi Bich Ngoc Trinh ◽  
Thang Truong ◽  
Tam Nguyen ◽  
Xuan Dang Vu ◽  
Anh Dao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Pcr Assay

scholarly journals Rapid Extraction and Detection of African Swine Fever Virus DNA Based on Isothermal Recombinase Polymerase Amplification Assay

Viruses ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1731
Author(s):  
Arianna Ceruti ◽  
Rea Maja Kobialka ◽  
Judah Ssekitoleko ◽  
Julius Boniface Okuni ◽  
Sandra Blome ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Dna Isolation ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Recombinase Polymerase Amplification ◽  
Reference Laboratory ◽  
Analytical Sensitivity ◽  
Lysis Buffer

African swine fever virus (ASFV) is the causative agent of a deadly disease in pigs and is spread rapidly across borders. Samples collected from suspected cases must be sent to the reference laboratory for diagnosis using polymerase chain reaction (PCR). In this study, we aimed to develop a simple DNA isolation step and real-time recombinase polymerase amplification (RPA) assay for rapid detection of ASFV. RPA assay based on the p72 encoding B646L gene of ASFV was established. The assays limit of detection and cross-reactivity were investigated. Diagnostic performance was examined using 73 blood and serum samples. Two extraction approaches were tested: silica-column-based extraction method and simple non-purification DNA isolation (lysis buffer and heating, 70 °C for 20 min). All results were compared with well-established real-time PCR. In a field deployment during a disease outbreak event in Uganda, 20 whole blood samples were tested. The assay’s analytical sensitivity was 3.5 DNA copies of molecular standard per µL as determined by probit analysis on eight independent assay runs. The ASFV RPA assay only detected ASFV genotypes. Compared to real-time PCR, RPA diagnostic sensitivity and specificity were 100%. Using the heating/lysis buffer extraction procedure, ASFV-RPA revealed better tolerance to inhibitors than real-time PCR (97% and 38% positivity rate, respectively). In Uganda, infected animals were identified before the appearance of fever. The ASFV-RPA assay is shown to be as sensitive and specific as real-time PCR. Moreover, the combination of the simple extraction protocol allows its use at the point of need to improve control measures.

Development of the test kit for detection of African swine fever virus by Real-Time PCR

2021 ◽  
Vol 24 (11) ◽  
pp. 17-22
Author(s):  
N.I. Salnikov ◽  
◽  
M.E. Senina ◽  
A.S. Preobrazhenskaya ◽  
Z.S. Devrishova ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus ◽  
Test Kit

scholarly journals Evaluation of an Automatic Insulated Isothermal PCR System for Rapid and Reliable Field-Deployable Detection of African Swine Fever Virus

2021 ◽  
Author(s):  
TRAN THI HUYEN NGA ◽  
LE THI NHI-CONG ◽  
PHAM BANG PHUONG ◽  
LUU VAN QUYNH ◽  
Viet-Linh Nguyen
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
High Specificity ◽  
Performance Comparison ◽  
Fever Virus ◽  
Analytical Sensitivity ◽  
Specificity Test ◽  
Insulated Isothermal Pcr

Abstract In the present study, we evaluate an automatic sample-to-answer insulated isothermal PCR system for rapid and reliable field-deployable detection of African swine fever virus in comparison to that of OIE recommended real-time PCR counterparts with samples collected in Vietnam. For analytical sensitivity, the system could detect ASFV up to a dilution of 106 whereas the real time PCR systems could detect up to a dilution of 105 to 106. For specificity test, the system showed high specificity to ASFV in compare to different other types of swine pathogens: PRRSV, FMDV, PCV2, CSFV. The diagnostic performance comparison on 6 different types of samples showed 97.3% to 100% agreement with reference real-time PCR. The results of this study indicated that POCKIT Central-based insulated isothermal PCR system is a rapid, reliable and sample-flexible method for effective detection of ASFV.

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