|
KMID : 0384119930130030437
|
|
Korean Journal of Clinical Pathology
1993 Volume.13 No. 3 p.437 ~ p.444
|
|
|
Optimal Condition of Polymerase Chain Reaction for the Detection of Mycobacterium tuberculosis
|
|
김한길
은상진/송경은/서장수/이원길/김재식/최성만
|
|
|
Abstract
|
|
|
Recently developed polymerase chain reaction(PCR) technology has been employed to Mycobacterium tuberculosis-specific DNA fragment. However, most studies concerned about the detection of M. tuberculosis in clinical specimen by PCR and few studies
have
compared DNA isolation methods, different kinds of polymerase, primer and thermocyeler. In this study, for the purpose of performing more effective PCR, we have compared two DNA isolation methods, 3 polymerases, 2 sets of primers and 2 kinds of
thermocycler. We evaluated two DNA isolation methods, and proteinase-K method was more sensitive than bead-beating method. In comparision of two sets of primers. P1 and P2 primers detecting 123-base pair fragment of IS6110 made from
Biosynthesis(U.S.A.)
and INSI and INS2 primers detecting 245-base pair fragment of IS986 showed equally sensitive result. Specificity of the primers was tested and INS1 and INS2 primers gave 245-base pair product from M. tuberculosis, but not from eleven other
bacterial
strains commonly found in the upper respiratory tract. Specificity of the P1 and P2 primers was not tested in this study. In comparision of polymerases, Taq DNA polymerasesR(Promega Co., U.S.A.) was more cost effective than AmpliTaqR(Perkin-Elmer
Cetus,
U.S.A.), but these two were equally sensitive and specific in the detection of M. tuberculosis-specific DNA. In comparision of thermocycler, Easy-CyclerR(Ericomp, U.S.A.) was more sensitive than water bath type.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|
|