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Application of immunoperoxidase staining to more rapid detection and identification of rubella virus isolates
Abstract:
Efforts were made to shorten the time required for detection of rubella virus in clinical materials through the use of immunoperoxidase (IP) staining. Comparative studies were performed in which specimens were inoculated in parallel into BHK-21 hamster kidney cells, which were examined by IP staining at 5 days, and into BK-13 and BS-C-1 cells, which were examined in two ways, viz., by subpassage at 7 days into BHK-21 cells and IP staining 3 days later and by subpassage at 7 days into BS-C-1 cells followed by interference testing and immunofluorescence (IF) staining on positive materials (standard method). Direct inoculation into BHK-21 cells with IP staining at 5 days permitted detection and identification of 59% of the 63 positive specimens. Toxicity of some specimens preserved with sorbitol and of certain tissue specimens reduced the number of satisfactory examinations which could be performed in this system. Virus detection and identification by IP staining on subpassaged RK-13 and BS-C-1 materials, requiring a total of 18 days, was comparable to the longer interference-IF method, requiring 17 days. Results obtained by IP staining and interference-IF showed 98% correlation on RK-13 materials and 97% correlation on BS-C-1 materials. IP staining on inoculated BHK-21 cells can be a useful method for rapid identification of a relatively high proportion of rubella-positive specimens, particularly if sorbitol-preserved specimens are avoided, and IP staining on subpassaged RK-13 and BS-C-1 materials is a highly satisfactory alternative to the longer interference-IF method.
Insights
Immunoperoxidase (IP) staining offers a faster method for rubella virus detection in clinical samples. This technique provides a reliable alternative to traditional methods, improving diagnostic efficiency.
Area of Science:
- Virology
- Immunology
- Diagnostic Microbiology
Background:
- Rubella virus detection is crucial for timely diagnosis and management.
- Traditional methods for rubella virus identification can be time-consuming.
- There is a need for rapid and accurate diagnostic techniques.
Purpose of the Study:
- To evaluate the efficacy of immunoperoxidase (IP) staining for rapid rubella virus detection.
- To compare IP staining with the standard interference-immunofluorescence (IF) method.
- To assess the time efficiency of IP staining in clinical materials.
Main Methods:
- Comparative study of IP staining and interference-IF staining.
- Inoculation of clinical specimens into BHK-21, RK-13, and BS-C-1 cell lines.
- Direct IP staining at 5 days and subpassage with IP staining at 18 days.
- Standard method involved subpassage, interference testing, and IF staining (17 days).
Main Results:
- Direct IP staining on BHK-21 cells detected 59% of rubella-positive specimens.
- IP staining on subpassaged RK-13 and BS-C-1 cells showed high correlation (98% and 97%) with the standard method.
- The IP staining method on subpassaged cells (18 days) was comparable in accuracy to the interference-IF method (17 days).
- Specimen toxicity affected examination success rates.
Conclusions:
- IP staining on BHK-21 cells allows for rapid identification of a significant proportion of rubella-positive specimens.
- IP staining on subpassaged RK-13 and BS-C-1 cells is a viable and efficient alternative to the interference-IF method.
- Avoiding sorbitol-preserved specimens enhances the reliability of IP staining.