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

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 realtime polymerase chain reaction (PCR). Howev...

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Veröffentlicht in:Asian-Australasian journal of animal sciences. - 1998. - 33(2020), 10 vom: 11. Okt., Seite 1683-1690
1. Verfasser: Tran, Ha Thi Thanh (VerfasserIn)
Weitere Verfasser: 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
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2020
Zugriff auf das übergeordnete Werk:Asian-Australasian journal of animal sciences
Schlagworte:Journal Article African Swine Fever Conventional Polymerase Chain Reaction Molecular Diagnosis PAMs Cell Real-time Polymerase Chain Reaction Virus Isolation
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520 |a 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 realtime 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 
520 |a 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 
520 |a 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) 
520 |a 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 
650 4 |a Journal Article 
650 4 |a African Swine Fever 
650 4 |a Conventional Polymerase Chain Reaction 
650 4 |a Molecular Diagnosis 
650 4 |a PAMs Cell 
650 4 |a Real-time Polymerase Chain Reaction 
650 4 |a Virus Isolation 
700 1 |a Dang, Anh Kieu  |e verfasserin  |4 aut 
700 1 |a Ly, Duc Viet  |e verfasserin  |4 aut 
700 1 |a Vu, Hao Thi  |e verfasserin  |4 aut 
700 1 |a Hoang, Tuan Van  |e verfasserin  |4 aut 
700 1 |a Nguyen, Chinh Thi  |e verfasserin  |4 aut 
700 1 |a Chu, Nhu Thi  |e verfasserin  |4 aut 
700 1 |a Nguyen, Vinh The  |e verfasserin  |4 aut 
700 1 |a Nguyen, Huyen Thi  |e verfasserin  |4 aut 
700 1 |a Truong, Anh Duc  |e verfasserin  |4 aut 
700 1 |a Pham, Ngoc Thi  |e verfasserin  |4 aut 
700 1 |a Dang, Hoang Vu  |e verfasserin  |4 aut 
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