Improving sensitivity of single tube nested PCR to detect fastidious microorganisms.

ABSTRACT: Single Tube Nested PCR (ST-nPCR) is of value to clinical laboratories with limited settings for the detection of fastidious microorganisms. The detection sensitivity of ST-nPCR is dependent on ensuring minimal leftovers of outer primers during the second round of the reaction. In this work, we investigated various approaches to optimize the performance of outer primers, including decreasing outer primer concentrations; using antisense oligonucleotides to block outer primers; using chemically modified inner primers; and using Q5 Taq polymerase that lacks 5'-3' exonuclease and strand displacement capabilities. These solutions were tested on C. abortus and C. psittaci, which are both fastidious intracellular bacteria that are difficult to diagnose. The best obtained result was b

SUBMITTER: Shatleh-Rantisi D 

PROVIDER: S-EPMC7002839 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature