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2.
Mem Inst Oswaldo Cruz ; 101(6): 635-8, 2006 Sep.
Article in English | MEDLINE | ID: mdl-17072475

ABSTRACT

The bovine papillomavirus type 2 (BPV-2) involvement in the aetiology of chronic enzootic haematuria associated to bracken fern ingestion has been suggested for a long time. However, a few reports have shown the presence of the BPV-2 in urinary bladder tumors of cattle. The aim of this study was to investigate the presence of the BPV-2 infection in the urinary bladder of cattle with chronic enzootic haematuria in Brazilian cattle herds. Sixty-two urinary bladders were collected from adult cattle in beef herds from the north region of the state of Paraná, Brazil. According to clinical and pathological finds the specimens were distributed in three groups: the group A was constituted by 22 urinary bladders with macroscopic lesions collected at necropsy of cattle with clinical signs of chronic enzootic haematuria; the group B by 30 urinary bladders with macroscopic lesions collected in a slaughterhouse of cows coming from bracken fern-endemic geographical region; and the group C (control) by 10 urinary bladders without macroscopic lesions collected from asymptomatic cattle in a bracken fern-free geographical region. By a semi-nested polymerase chain reaction (PCR) assay, with an internal control, a fragment of the BPV-2 L1 gene with 386 bp length was amplified in 36 (58%) urinary bladder. The rate of BPV-2 positive urinary bladders was 50% (11/22) for group A, 80% (24/30) for group B, and 10% (1/10) for group C (control). The rate of the positive results found in groups A and B that included urinary bladder samples with macroscopic lesions was 67% (35/52) and the detection of the BPV-2 in both groups was significantly higher (P < 0.05) than in the control group. RFLP with Rsa I and Hae III enzymes evaluated the specificity of the BPV-2 amplicons. The PCR internal control that amplified a 626 bp fragment of the ND5 gene of the bovine mitochondrial genome was amplified in all analyzed samples and excluded false-negatives or invalid results in the semi-nested PCR. These results suggest the BPV-2 involvement in the chronic enzootic haematuria aetiology and open the perspective of the development of new strategies for the control of this disease that is the major cause of economical losses in beef herds from many Brazilian geographical regions.


Subject(s)
Bovine papillomavirus 1/isolation & purification , Cattle Diseases/virology , Hematuria/veterinary , Papillomavirus Infections/veterinary , Urinary Bladder/virology , Animals , Bovine papillomavirus 1/genetics , Cattle , Chronic Disease , Hematuria/virology , Papillomavirus Infections/virology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length
3.
Mem. Inst. Oswaldo Cruz ; 101(6): 635-638, Sept. 2006. ilus, tab
Article in English | LILACS | ID: lil-437056

ABSTRACT

The bovine papillomavirus type 2 (BPV-2) involvement in the aetiology of chronic enzootic haematuria associated to bracken fern ingestion has been suggested for a long time. However, a few reports have shown the presence of the BPV-2 in urinary bladder tumors of cattle. The aim of this study was to investigate the presence of the BPV-2 infection in the urinary bladder of cattle with chronic enzootic haematuria in Brazilian cattle herds. Sixty-two urinary bladders were collected from adult cattle in beef herds from the north region of the state of Paraná, Brazil. According to clinical and pathological finds the specimens were distributed in three groups: the group A was constituted by 22 urinary bladders with macroscopic lesions collected at necropsy of cattle with clinical signs of chronic enzootic haematuria; the group B by 30 urinary bladders with macroscopic lesions collected in a slaughterhouse of cows coming from bracken fern-endemic geographical region; and the group C (control) by 10 urinary bladders without macroscopic lesions collected from asymptomatic cattle in a bracken fern-free geographical region. By a semi-nested polymerase chain reaction (PCR) assay, with an internal control, a fragment of the BPV-2 L1 gene with 386 bp length was amplified in 36 (58 percent) urinary bladder. The rate of BPV-2 positive urinary bladders was 50 percent (11/22) for group A, 80 percent (24/30) for group B, and 10 percent (1/10) for group C (control). The rate of the positive results found in groups A and B that included urinary bladder samples with macroscopic lesions was 67 percent (35/52) and the detection of the BPV-2 in both groups was significantly higher (P < 0.05) than in the control group. RFLP with Rsa I and Hae III enzymes evaluated the specificity of the BPV-2 amplicons. The PCR internal control that amplified a 626 bp fragment of the ND5 gene of the bovine mitochondrial genome was amplified in all analyzed samples and excluded false-negatives or invalid results in the semi-nested PCR...


Subject(s)
Animals , Cattle , Bovine papillomavirus 1/isolation & purification , Cattle Diseases/virology , Hematuria/veterinary , Papillomavirus Infections/veterinary , Urinary Bladder/virology , Bovine papillomavirus 1/genetics , Chronic Disease , Hematuria/virology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Papillomavirus Infections/virology
4.
J Clin Virol ; 27(3): 270-5, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12878091

ABSTRACT

BACKGROUND: Adenovirus-associated hemorrhagic cystitis (HC) has become a recognized sequel of immunosuppression. The diagnosis of viral infection is usually determined by viral cultures. OBJECTIVES: Analysis of different diagnostic methods for adenovirus (AdV) detection in bone marrow transplant patients with hemorrhagic cystitis. STUDY DESIGN: We describe a prospective study for AdV detection in the urine of patients with hematuria in the first 100 days after bone marrow transplant (BMT), comparing different laboratory techniques, PCR, enzyme immunoassay (EIA) and conventional culture. RESULTS: A total of 143 urine samples were analyzed, 75 collected in the pre-transplant period with and without hematuria and 68 post-transplant, only with microscopic or macroscopic hematuria. After BMT, hematuria occurred in 38.9% of patients, being more frequent in unrelated donor transplants. AdV was isolated in one pre-transplant patient without symptoms and in three post-transplant patients with HC grades 3 and 4 (severe), who were in month 2 or 3 post-transplant. Compared to culture as the gold standard, the accuracy, specificity and sensitivity of EIA were 95, 30 and 100% and for PCR were 63, 100 and 60%, respectively. CONCLUSIONS: We concluded that despite technical difficulties and the long time that elapsed before results were obtained, cell culture still remains the best method for adenovirus detection in the urine of patients with hemorrhagic cystitis.


Subject(s)
Adenovirus Infections, Human/diagnosis , Adenoviruses, Human/isolation & purification , Bone Marrow Transplantation/adverse effects , Cystitis/virology , Hematuria/virology , Adenovirus Infections, Human/virology , Adenoviruses, Human/genetics , Adolescent , Adult , Cell Line , Child , Child, Preschool , Female , Humans , Immunoenzyme Techniques , Male , Middle Aged , Polymerase Chain Reaction , Sensitivity and Specificity , Urine/virology , Virus Cultivation
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