|
NISCAIR ONLINE PERIODICALS REPOSITORY (NOPR) >
NISCAIR PUBLICATIONS >
Research Journals >
Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.49 [2011] >
IJEB Vol.49(04) [April 2011] >
| Title: | A sensitive and specific ES-31 antigen detection based fluorometric assay for confirmation of Mycobacterium tuberculosis in cell culture |
| Authors: | Majumdar, Anindita
Wankhade, Gauri
Kamble, Pranita D
Joshi, Deepti
Harinath, B C |
| Keywords: | Diagnostic tests
ES-31 antigen
Immunofluorescent staining
Mycobacterium tuberculosis |
| Issue Date: | Apr-2011 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | Confirmation of presence of M. tuberculosis bacilli on microscopic examination is very
important in diagnosis of tuberculosis. The present study was undertaken to
find the usefulness of mycobacterial ES-31 serine protease as a marker to
detect tuberculosis bacilli using fluorescein isothiocyanate conjugated
anti-ES-31 serine protease antibody. This immunofluorescence method was
compared with Ziehl-Neelsen and auramine-O staining methods for detection of
tuberculosis bacilli. Slides were prepared for each serially diluted
tuberculosis H37Ra bacilli (1×107 bacilli/ml to 5 bacilli/ml).
Slides for each dilution group were stained by ZN method, auramine-O and
immunostaining methods using fluorescein isothiocyanate conjugated anti-ES-31
serine protease antibody. ZN staining method showed efficacy for detection of M. tuberculosis H37Ra upto
1×104 bacilli/ml while auramine-O method showed upto 1×102
bacilli/ml. The presence of bacilli was indicated by green fluorescence on
immunostaining using anti-ES-31 antibody conjugate and this method was
effective upto 10 bacilli/ml. The slides which were negative for ZN (1×103
cells/ml) and auramine-O (100 cells/ml) method showed positivity on restaining
with immunofluorescent staining method. The results of this preliminary study
showed that immunofluorescent staining method using specific anti-ES-31
antibody conjugate was more sensitive for detection of tuberculosis bacilli
than ZN and auramine-O methods in samples of laboratory strain. The utility of
this method will be studied further in clinical specimens. |
| Page(s): | 304-306 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.49(04) [April 2011]
|
|
