Analytic performance and contamination control methods of a ligase chain reaction DNA amplification assay for detection of Chlamydia trachomatis in urogenital specimens

1996 ◽  
Vol 24 (2) ◽  
pp. 71-76 ◽  
Author(s):  
Hsiang-Yun Y. Hu ◽  
John D. Burczak ◽  
Gregor W. Leckie ◽  
Keith A. Ray ◽  
Sharon Muldoon ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Dna Amplification ◽  
Control Methods ◽  
Chain Reaction ◽  
Contamination Control ◽  
Ligase Chain Reaction ◽  
Amplification Assay

scholarly journals Method for Reduction of Inhibition in aMycobacterium tuberculosis-Specific Ligase Chain Reaction DNA Amplification Assay

1998 ◽  
Vol 36 (3) ◽  
pp. 764-767 ◽  
Author(s):  
Gregor W. Leckie ◽  
Dwight D. Erickson ◽  
Qizhi He ◽  
Ingrid E. Facey ◽  
Bor-Chian Lin ◽  
...  
Keyword(s):  
Solubility Product ◽  
Dna Amplification ◽  
Acidic Ph ◽  
Major Contributor ◽  
Chain Reaction ◽  
Ligase Chain Reaction ◽  
Phosphate Ions ◽  
Amplification Assay ◽  
Culture Positive ◽  
Respiratory Specimens

The present study describes the identification of inhibitors of aMycobacterium tuberculosis-specific gap ligase chain reaction (LCR) DNA amplification assay as well as a method for their removal. A major contributor to inhibition was deduced to be a calcium phosphate precipitate, CaHPO4. The precipitate forms duringN-acetyl-l-cysteine–sodium hydroxide (NALC-NaOH) decontamination, digestion, and concentration of respiratory specimens. The solubility product of CaHPO4 precipitate at pH 7.8, the pH at which gap LCR is optimized, indicates that the precipitate releases an amount of phosphate ions sufficient to inhibit amplification. A method for removal of the precipitate was identified. The precipitate is dissociated by exposing it to a mildly acidic (pH 4.1) buffer during the first of two centrifugation steps; the inhibitory phosphate ions are removed by the centrifugation steps. When 100 NALC-NaOH respiratory sediments were tested by gap LCR, none of the sediments were inhibitory when the acidic buffer was used while 24 samples were inhibitory when TE buffer, pH 7.8, was used. In another study, when the acidic buffer wash was applied to 1,440 NALC-NaOH respiratory sediments, only 10 sediments were found to be inhibitory. None of the inhibited sediments were culture positive for M. tuberculosis. This work demonstrates that when inhibition mechanisms are identified, relatively simple protocols can be used to obtain low inhibition rates and to allow the use of larger volume equivalents in amplification reactions.

scholarly journals Comparison of the ligase chain reaction with cell culture for the diagnosis of Chlamydia trachomatis infection in women.

1996 ◽  
Vol 49 (2) ◽  
pp. 116-119 ◽  
Author(s):  
G L Ridgway ◽  
G Mumtaz ◽  
A J Robinson ◽  
M Franchini ◽  
C Carder ◽  
...  
Keyword(s):  
Cell Culture ◽  
Chlamydia Trachomatis ◽  
Chain Reaction ◽  
Ligase Chain Reaction ◽  
Chlamydia Trachomatis Infection

Amplification of Chlamydia trachomatis DNA by ligase chain reaction.

1993 ◽  
Vol 31 (3) ◽  
pp. 729-731 ◽  
Author(s):  
B J Dille ◽  
C C Butzen ◽  
L G Birkenmeyer
Keyword(s):  
Chlamydia Trachomatis ◽  
Chain Reaction ◽  
Ligase Chain Reaction

scholarly journals Inhibition of amplification of Chlamydia trachomatis plasmid DNA by the ligase chain reaction associated with female urines

1998 ◽  
Vol 4 (7) ◽  
pp. 397-400 ◽  
Author(s):  
Max Chernesky ◽  
Sylvia Chong ◽  
Dan Jang ◽  
Kathy Luinstra ◽  
Michael Faught ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Plasmid Dna ◽  
Chain Reaction ◽  
Ligase Chain Reaction
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