Experimental infection with the Paderborn isolate of classical swine fever virus in 10-week-old pigs:: determination of viral replication kinetics by quantitative RT-PCR, virus isolation and antigen ELISA

被引:61
作者
Uttenthal, Å [1 ]
Storgaard, T [1 ]
Oleksiewicz, MB [1 ]
de Stricker, K [1 ]
机构
[1] Danish Vet Inst, DK-4771 Lindholm, Kalvehave, Denmark
关键词
classical swine fever virus; Paderborn isolate; hog cholera virus; quantitative RT-PCR; experimental infection; diagnosis; meat juice;
D O I
10.1016/S0378-1135(02)00364-4
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We performed experimental infection in 10-week-old pigs with the Paderborn isolate of classical swine fever virus (CSFV). Despite being epidemiologically linked to the major CSFV outbreak in The Netherlands in 1997, the in vivo replication kinetics of this isolate have to our knowledge not been described in detail previously. We found that oronasal infection with 10(4.7) TCID50 produced mortality in three out of five pigs after 29-31 days, and severe clinical symptoms in one out of five pigs, while one out of five pigs exhibited no clinical symptoms. At this infection dose, pigs had viral RNA (monitored by quantitative reverse transcription (RT)-PCR) in serum as soon as 2 days post-infection, and excretion of infectious virus (monitored by sentinel pigs) appeared to be virtually concomitant with viremia onset. While virus RNA was cleared from the serum of most pigs after 1-2 weeks, some pigs had viral RNA in serum for more than 30 days, and exhibited only mild clinical symptoms. We observed an excellent correlation between clinical symptoms and viral RNA loads in serum, while serum antibody levels were low. Clinically affected pigs had up to 1000-fold higher serum viral RNA loads than did pigs without clinical symptoms. At this level of infection, and this age group, the Paderborn isolate exhibited a strikingly wide range of replication patterns, which might be relevant to the spread of the virus through susceptible pig populations, and the severity of the 1997-1998 outbreak. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:197 / 212
页数:16
相关论文
共 26 条
[1]   RAPID AND SIMPLE METHOD FOR PURIFICATION OF NUCLEIC-ACIDS [J].
BOOM, R ;
SOL, CJA ;
SALIMANS, MMM ;
JANSEN, CL ;
WERTHEIMVANDILLEN, PME ;
VANDERNOORDAA, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (03) :495-503
[2]   SPECIFIC SEQUENCE AMPLIFICATION OF BOVINE VIRUS DIARRHEA VIRUS (BVDV) AND HOG-CHOLERA VIRUS AND SEQUENCING OF BVDV NUCLEIC-ACID [J].
BOYE, M ;
KAMSTRUP, S ;
DALSGAARD, K .
VETERINARY MICROBIOLOGY, 1991, 29 (01) :1-13
[3]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[4]  
DAHLE J, 1995, WIEN TIERARZTL MONAT, V82, P232
[5]   Classical swine fever (CSF) marker vaccine - Trial II. Challenge study in pregnant sows [J].
Depner, KR ;
Bouma, A ;
Koenen, F ;
Klinkenberg, D ;
Lange, E ;
de Smit, H ;
Vanderhallen, H .
VETERINARY MICROBIOLOGY, 2001, 83 (02) :107-120
[6]   An experimental infection with classical swine fever in E2 subunit marker-vaccine vaccinated and in non-vaccinated pigs [J].
Dewulf, J ;
Laevens, H ;
Koenen, F ;
Vanderhallen, H ;
Mintiens, K ;
Deluyker, H ;
de Kruif, A .
VACCINE, 2000, 19 (4-5) :475-482
[7]   The classical swine fever epidemic 1997-1998 in the Netherlands: descriptive epidemiology [J].
Elbers, ARW ;
Stegeman, A ;
Moser, H ;
Ekker, HM ;
Smak, JA ;
Pluimers, FH .
PREVENTIVE VETERINARY MEDICINE, 1999, 42 (3-4) :157-184
[8]   Classical swine fever (CSF) marker vaccine - Trial III. Evaluation of discriminatory ELISAs [J].
Floegel-Niesmann, G .
VETERINARY MICROBIOLOGY, 2001, 83 (02) :121-136
[9]   Application of a computer program for genetic typing of classical swine fever virus isolates from Germany [J].
Greiser-Wilke, I ;
Depner, K ;
Fritzemeier, J ;
Haas, L ;
Moennig, V .
JOURNAL OF VIROLOGICAL METHODS, 1998, 75 (02) :141-150
[10]   Molecular epidemiology of a large classical swine fever epidemic in the European Union in 1997-1998 [J].
Greiser-Wilke, I ;
Fritzemeier, J ;
Koenen, F ;
Vanderhallen, H ;
Rutili, D ;
De Mia, GM ;
Romero, L ;
Rosell, R ;
Sanchez-Vizcaino, JM ;
San Gabriel, A .
VETERINARY MICROBIOLOGY, 2000, 77 (1-2) :17-27