The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples
Primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala
Апстракт
The objective of this study was the application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. Twenty samples of bovine nasal swabs were separately inoculated in Vero cells. Laboratory strain TN 41 of bovine herpesvirus 1 served as a control in the experiment. The cytopathic effects were recovered in the cell line previously inoculated with a sample of bovine nasal swab, as well as in cells inoculated with the laboratory strain TN 41 of the bovine herpesvirus 1 after a period of incubation of 24h, 36h and 48h. Identification of the isolated strains of the virus was done by the virus - neutralization test with specific immune sera against BHV 1, with a titre of 1:16. Comparative analysis of DNA fragments of the laboratory strain of BHV1 and isolated strain obtained by polymerase chain reaction with primers for viral gB glycoprotein and thymidine-kinase coding region, confirmed that the isolated strain of the virus belongs to bovine herpesvirus 1 (...BHV 1).
Cilj nàših istraživanja je bio primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala. Dvadeset uzoraka nosnih briseva goveda pojedinačno je inokulisano u ćelijsku liniju Vero. Referentni soj TN 41 virusa BHV 1 služio je kao pozitivna kontrola u ogledu. Posle 24h, 36h i 48h od inokulacije, utvrđena je pojava citopatogenog efekta u ćelijskoj liniji inokulisanoj uzorkom poreklom od jednog nosnog brisa goveda. Iste promene su ustanovljene i posle inokulacije referentnog soja TN 41 virusa BHV 1 u ćelijsku liniju Vero. Identifikacija izolovanog soja virusa poreklom iz uzorka nosnog brisa goveda, vršena je primenom testa virus - neutralizacije uz korišćenje specifičnog imunog seruma protiv virusa BHV 1, titra od 1:16. Uporednom analizom DNK fragmenata referentnog soja virusa BHV 1 i izolovanog soja virusa, dobijenih primenom lančane reakcije polimeraze uz korišćenje dva para prajmera koji su amplifikovali gene na molekulu DNK koji k...odiraju sintezu glikoproteina B spoljašnjeg omotača virusa BHV 1 i timidin - kinaze, potvrđena je pripadnost izolovanog soja virusa goveđem herpesvirusu 1.
Кључне речи:
bovine herpesvirus 1 (BHV 1) / Vero cells / virus -neutralization test (VN-test) / virus isolation / polymerase chain reaction(PCR)Извор:
Acta Veterinaria-Beograd, 2010, 60, 1, 39-48Издавач:
- Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
Финансирање / пројекти:
- Развој и примена ланчане реакције полимеразе (PCR) у брзој дијагностици респираторне инфекције говеда изазване говеђим херпесвирусом 1 (RS-MESTD-MPN2006-2010-20150)
- Биолошке последице генетичке варијабилности вируса (RS-MESTD-MPN2006-2010-145047)
DOI: 10.2298/AVB1001039N
ISSN: 0567-8315
WoS: 000276046300005
Scopus: 2-s2.0-77950650435
Колекције
Институција/група
Fakultet veterinarske medicineTY - JOUR AU - Nišavić, Jakov AU - Milić, Nenad AU - Knežević, Aleksandra AU - Jovanović, Tanja PY - 2010 UR - https://vet-erinar.vet.bg.ac.rs/handle/123456789/662 AB - The objective of this study was the application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. Twenty samples of bovine nasal swabs were separately inoculated in Vero cells. Laboratory strain TN 41 of bovine herpesvirus 1 served as a control in the experiment. The cytopathic effects were recovered in the cell line previously inoculated with a sample of bovine nasal swab, as well as in cells inoculated with the laboratory strain TN 41 of the bovine herpesvirus 1 after a period of incubation of 24h, 36h and 48h. Identification of the isolated strains of the virus was done by the virus - neutralization test with specific immune sera against BHV 1, with a titre of 1:16. Comparative analysis of DNA fragments of the laboratory strain of BHV1 and isolated strain obtained by polymerase chain reaction with primers for viral gB glycoprotein and thymidine-kinase coding region, confirmed that the isolated strain of the virus belongs to bovine herpesvirus 1 (BHV 1). AB - Cilj nàših istraživanja je bio primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala. Dvadeset uzoraka nosnih briseva goveda pojedinačno je inokulisano u ćelijsku liniju Vero. Referentni soj TN 41 virusa BHV 1 služio je kao pozitivna kontrola u ogledu. Posle 24h, 36h i 48h od inokulacije, utvrđena je pojava citopatogenog efekta u ćelijskoj liniji inokulisanoj uzorkom poreklom od jednog nosnog brisa goveda. Iste promene su ustanovljene i posle inokulacije referentnog soja TN 41 virusa BHV 1 u ćelijsku liniju Vero. Identifikacija izolovanog soja virusa poreklom iz uzorka nosnog brisa goveda, vršena je primenom testa virus - neutralizacije uz korišćenje specifičnog imunog seruma protiv virusa BHV 1, titra od 1:16. Uporednom analizom DNK fragmenata referentnog soja virusa BHV 1 i izolovanog soja virusa, dobijenih primenom lančane reakcije polimeraze uz korišćenje dva para prajmera koji su amplifikovali gene na molekulu DNK koji kodiraju sintezu glikoproteina B spoljašnjeg omotača virusa BHV 1 i timidin - kinaze, potvrđena je pripadnost izolovanog soja virusa goveđem herpesvirusu 1. PB - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd T2 - Acta Veterinaria-Beograd T1 - The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples T1 - Primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala VL - 60 IS - 1 SP - 39 EP - 48 DO - 10.2298/AVB1001039N ER -
@article{ author = "Nišavić, Jakov and Milić, Nenad and Knežević, Aleksandra and Jovanović, Tanja", year = "2010", abstract = "The objective of this study was the application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. Twenty samples of bovine nasal swabs were separately inoculated in Vero cells. Laboratory strain TN 41 of bovine herpesvirus 1 served as a control in the experiment. The cytopathic effects were recovered in the cell line previously inoculated with a sample of bovine nasal swab, as well as in cells inoculated with the laboratory strain TN 41 of the bovine herpesvirus 1 after a period of incubation of 24h, 36h and 48h. Identification of the isolated strains of the virus was done by the virus - neutralization test with specific immune sera against BHV 1, with a titre of 1:16. Comparative analysis of DNA fragments of the laboratory strain of BHV1 and isolated strain obtained by polymerase chain reaction with primers for viral gB glycoprotein and thymidine-kinase coding region, confirmed that the isolated strain of the virus belongs to bovine herpesvirus 1 (BHV 1)., Cilj nàših istraživanja je bio primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala. Dvadeset uzoraka nosnih briseva goveda pojedinačno je inokulisano u ćelijsku liniju Vero. Referentni soj TN 41 virusa BHV 1 služio je kao pozitivna kontrola u ogledu. Posle 24h, 36h i 48h od inokulacije, utvrđena je pojava citopatogenog efekta u ćelijskoj liniji inokulisanoj uzorkom poreklom od jednog nosnog brisa goveda. Iste promene su ustanovljene i posle inokulacije referentnog soja TN 41 virusa BHV 1 u ćelijsku liniju Vero. Identifikacija izolovanog soja virusa poreklom iz uzorka nosnog brisa goveda, vršena je primenom testa virus - neutralizacije uz korišćenje specifičnog imunog seruma protiv virusa BHV 1, titra od 1:16. Uporednom analizom DNK fragmenata referentnog soja virusa BHV 1 i izolovanog soja virusa, dobijenih primenom lančane reakcije polimeraze uz korišćenje dva para prajmera koji su amplifikovali gene na molekulu DNK koji kodiraju sintezu glikoproteina B spoljašnjeg omotača virusa BHV 1 i timidin - kinaze, potvrđena je pripadnost izolovanog soja virusa goveđem herpesvirusu 1.", publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd", journal = "Acta Veterinaria-Beograd", title = "The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples, Primena lančane reakcije polimeraze u dokazivanju prisustva goveđeg herpesvirusa 1 u kliničkim uzorcima materijala", volume = "60", number = "1", pages = "39-48", doi = "10.2298/AVB1001039N" }
Nišavić, J., Milić, N., Knežević, A.,& Jovanović, T.. (2010). The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. in Acta Veterinaria-Beograd Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 60(1), 39-48. https://doi.org/10.2298/AVB1001039N
Nišavić J, Milić N, Knežević A, Jovanović T. The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples. in Acta Veterinaria-Beograd. 2010;60(1):39-48. doi:10.2298/AVB1001039N .
Nišavić, Jakov, Milić, Nenad, Knežević, Aleksandra, Jovanović, Tanja, "The application of polymerase chain reaction in detection of bovine herpesvirus 1 in clinical samples" in Acta Veterinaria-Beograd, 60, no. 1 (2010):39-48, https://doi.org/10.2298/AVB1001039N . .