RESUMEN
Two clinically healthy mature Pakistani Bos indicus × Bos taurus cattle were genotyped as homozygous affected for the lethal immunodeficiency disorder bovine leukocyte adhesion deficiency (BLAD) using previously described PCR-RFLP based DNA tests which was confirmed by sequencing. Sequencing of Bos taurus and B. indicus × B. taurus genomic DNA surrounding the disease causing mutation (c.383A > G) in the ITGB2 gene identified numerous variations in exonic and intronic regions within and between species, including substantial variation in primer annealing sites for three PCR-RFLP tests for one of the B. indicus allelic variants. These variations in the primer annealing sites resulted in a null allele in the DNA tests causing the misdiagnosis of some heterozygous B. taurus × B. indicus cattle to be classified as homozygous affected. New primers were designed and a modified test was developed which simultaneously identified the disease mutation and the Pakistani B. indicus allelic variant associated with the null allele in the previous test.
Asunto(s)
Enfermedades de los Bovinos/genética , Pruebas Genéticas , Síndrome de Deficiencia de Adhesión del Leucocito/veterinaria , Alelos , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Animales , Secuencia de Bases , Antígenos CD18/genética , Bovinos , Enfermedades de los Bovinos/diagnóstico , Estudios de Asociación Genética , Síndrome de Deficiencia de Adhesión del Leucocito/diagnóstico , Síndrome de Deficiencia de Adhesión del Leucocito/genética , Masculino , Técnicas de Diagnóstico Molecular , Datos de Secuencia Molecular , Pakistán , Análisis de Secuencia de ADNRESUMEN
Hepatitis C virus (HCV) is normally present in the blood of infected patients; however, it can also be present in some other body fluids. Therefore, in this study, a concurrent presence of HCV-RNA was investigated in oral fluid and urine of 80 Pakistani chronic HCV patients. HCV-RNA was detected in 31 (38.8%) oral fluid and 10 (12.5%) urine samples using RT-PCR in all 80 of the patients whose sera tested positive for HCV-RNA. From this study, it is concluded that, in addition to the blood, HCV RNA can also be found in oral secretions as well as urine of chronic HCV patients.
Asunto(s)
Hepacivirus/aislamiento & purificación , Hepatitis C Crónica/virología , ARN Viral/análisis , Adolescente , Adulto , Femenino , Hepacivirus/genética , Hepatitis C Crónica/sangre , Hepatitis C Crónica/orina , Humanos , Masculino , Persona de Mediana Edad , Pakistán , ARN Viral/sangre , ARN Viral/orina , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Saliva/virología , Orina/virologíaRESUMEN
Imatinib (Gleevec) is the effective therapy for BCR-ABL positive CML patients. Point mutations have been detected in ATP-binding domain of ABL gene which disturbs the binding of Gleevec to this target leading to resistance. Detection of mutations is helpful in clinical management of imatinib resistance. We established a very sensitive (ASO) PCR to detect mutations in an imatinib-resistant CML patient. Mutations C944T and T1052C were detected which cause complete partial imatinib resistance, respectively. This is the first report of multiple point mutations conferring primary imatinib resistance in same patient at the same time. Understanding the biological reasons of primary imatinib resistance is one of the emerging issues of pharmacogenomics and will be helpful in understanding primary resistance of molecularly-targeted cancer therapies. It will also be of great utilization in clinical management of imatinib resistance. Moreover, this ASO-PCR assay is very effective in detecting mutations related to imatinib resistance.