A single-step TaqMan® RT-qPCR was developed for the detection of bovine viral diarrhea virus type 1 (BVDV-1), an important pathogen of cattle worldwide. The assay was based on conserved 5'UTR sequences of Italian BVDV-1 isolates. In order to establish a diagnostic protocol which simplifies sample collection and processing, the assay was tested on a variety of biological specimens collected from persistently infected calves. The samples analyzed included PBMCs, plasma, dry blood, ear notch and hair bulb. Time and costs required for the analysis of each type of specimen were compared. The RT-qPCR, whose lower limit of detection was 100 copies of viral RNA (1 TCID50), correctly identified all PI animals, irrespective of the type of specimen. The highest copy numbers were obtained from the RNAs extracted from PBMCs, ear notches and hair bulbs. Hair bulb-supernatants directly used as a template allowed identification of all PI animals. In conclusion, based on time and cost evaluation, the most effective and efficient protocol was the one based on the direct analysis of hair bulb-supernatants, avoiding the RNA extraction step.
- MeSH
- bovinní diarea diagnóza virologie MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- RNA virová genetika MeSH
- skot MeSH
- Taq-polymerasa genetika metabolismus MeSH
- virus bovinní diarey 1 klasifikace genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- Geografické názvy
- Itálie MeSH
Enterocytozoon bieneusi known as a causative agent of opportunistic infections instigating diarrhoea in AIDS patients was identified also in a number of immunocompetent patients and in a wide range of animals, including cattle. In the present study we tested if the Bovine Viral Diarrhea Virus (BVDV), the most common pathogen underlying immunosuppressive Bovine Viral Diarrhoea (BVD), can enhance the occurrence of opportunistic infections with E. bieneusi in cattle. Six dairy farms were investigated using ELISA to detect antibodies against or antigens arising from BVDV in collected sera. A total of 240 individual faecal samples from four age groups were examined for the presence of E. bieneusi by nested PCR. Sequence analysis of six E. bieneusi positive samples revealed the presence of the genotype I of E. bieneusi, previously described in cattle. The hypothesis expecting higher prevalence of E. bieneusi in BVDV positive cattle herds was not confirmed in this study; however this is the first description about E. bieneusi in cattle in the Czech Republic.
- MeSH
- bovinní diarea komplikace mikrobiologie virologie MeSH
- ELISA veterinární MeSH
- Enterocytozoon * genetika MeSH
- feces mikrobiologie MeSH
- genotyp MeSH
- mikrosporidióza epidemiologie etiologie mikrobiologie veterinární virologie MeSH
- nemoci skotu epidemiologie mikrobiologie MeSH
- polymerázová řetězová reakce veterinární MeSH
- prevalence MeSH
- sekvenční analýza DNA veterinární MeSH
- skot parazitologie virologie MeSH
- věkové faktory MeSH
- viry bovinní diarey * MeSH
- zvířata MeSH
- Check Tag
- skot parazitologie virologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- bovinní diarea virologie MeSH
- finanční podpora výzkumu jako téma MeSH
- virus bovinní diarey 1 genetika MeSH
- virus bovinní diarey 2 genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Geografické názvy
- Argentina MeSH
- Brazílie MeSH
- Egypt MeSH
- Finsko MeSH
- Slovenská republika MeSH
- MeSH
- bovinní diarea virologie MeSH
- genom MeSH
- klasifikace MeSH
- sekvence nukleotidů MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- MeSH
- bovinní diarea patofyziologie virologie MeSH
- protilátky fyziologie krev MeSH
- skot MeSH
- Check Tag
- skot MeSH
- MeSH
- bovinní diarea patofyziologie virologie MeSH
- skot MeSH
- Check Tag
- skot MeSH
- MeSH
- bovinní diarea patofyziologie virologie MeSH
- protilátky fyziologie krev MeSH
- skot MeSH
- Check Tag
- skot MeSH
- MeSH
- bovinní diarea patofyziologie virologie MeSH
- skot MeSH
- Check Tag
- skot MeSH
