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The causal association of human papilloma virus (HPV) with cervical cancer has been supported by multiple lines of evidence. Therefore, in the case of dysplasia, the presence of HPV-DNA should be detected and its subtypes identified. This is important in the determination of the prognosis for cervical disease. We reported a study in which the localization and types of HPV in cervical diseases was identified by in situ hybridization using biotin-labelled DNA probes. Seven types (3 basic forms) of HPV were used as DNA probes. HPV types used were following: 6/11, 16/18, 31/33/35. In this study, we introduced 7 new types (3 forms) of probes for a total of 14 types (6 forms). The new probes introduced were 42/43/45, 45/56, 51/52. Using these probes, the rate of detection HPV according to types was examined. In addition, localization of HPV infection and its relationship with histopathological findings of cervical disease were evaluated. 1) Types 6/11 and 45/56 were found in the lesions with less important histological findings (CIN I). Types 16/18, 31/33/35, 51/52 were found in the all lesions. Type 42/43/44 were not detected in this study. 2) The addition of 7 new types (3 new forms) of probes resulted in a 17% increase (16 cases) in the rate of detection of HPV compared to our previous study. 3) Localization and distribution pattern of the HPV is not, as has been reported so far, dependent on HPV type but rather on the histological characteristics, such as the degree of dysplasia.
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Rotavirus infections in intestinal tissues of animals or in tissue culture cells were detected by the immunocytochemical unlabeled soluble enzyme peroxidase antiperoxidase method. Comparison of the immunofluorescence and peroxidase antiperoxidase immunological staining techniques revealed that the two methods are equally sensitive for detection of rotavirus-infected cells. The peroxidase antiperoxidase technique offers the advantages of negligible nonspecific staining reactions, the use of a standard light microscope, the production of permanent slides, and the conservation of immunological reagents. The ability to detect antigens in paraffin-embedded tissues enhances the usefulness of the peroxidase antiperoxidase test for both prospective and retrospective studies.
A comparison was made between a biuret (reference) method and a simple refractometric (test) method for measuring fibrinogen levels in 84 animal plasmas. Although the correlation between the two methods was high (4=0.90 P less than 0-001) there was considerable random variation in the refractometric results in relation to the biuret results. This was thought to be due in part to the fact that refractometric results could only be expressed in multiples of 2.4 g/litre. In spite of this limitation, the refractometric method, on the grounds of speen and simplicity, is considered to have worthwhile application for fibrinogen determinations in practice laboratory.
The diagnosis of genetic infections and cancerous diseases is carried out more and more often at a molecular level using Southern's technique which is based on the use of 32P-labelled DNA. In order to circumvent the risks and rapid decrease in radioactivity associated with these latter techniques, new colorigenic methods have been developed. In this work, we describe the use of dTTP analogues (digoxigenin-dUTP and biotin-dUTP) for the labelling of probes and detection of target DNA. Using digoxigenin-11-dUTP, 0.1 aM of a 561 bp target DNA was detected by using a modified Southern procedure. The reliability and the high sensitivity of such methods make them a good tool for DNA investigation in research as well as in testing laboratories.