| Application of polymerase chain reaction(PCR) to the diagnosis of tuberculosis. |
|
Ho Joong Kim, Young Whan Kim, Sung Koo Han, Young Soo Shim, Keun Youl Kim, Yong Chol Han |
|
Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea |
|
|
| Abstract |
Background
Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis.
Methods
To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB.1, .2) , and 123 base pair DNA, a part of IS6110 fragment , which multiple copies are known to exsist per one genome (the primers are Sal I.1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results.
Results
1) Using TB-l , -2 primers, PCR was positive in 73.1 % (19/ 26) of culture positive sputa, in 12.5% (1 / 8) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology.
2) Using Sal 1-1, -2 primers, PCR was positive in 94.1% (32 / 34) of culture positive sputa, in 23.1% (6/ 26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14 / 16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology.
Conclusions
PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method. |
| Key Words:
PCR, Mycobacterium tuberculosis, Sputum |
|
PDF Links
Full text via DOI
Download Citation
