Increased Sensitivity and Specificity of Borrelia burgdorferi 16S Ribosomal DNA,

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Increased Sensitivity and Specificity of Borrelia

burgdorferi 16S Ribosomal DNA,Detection.

 

Am J Clin Pathol. 2010 Apr;133(4):569-76.

 

Increased Sensitivity and Specificity of Borrelia burgdorferi 16S

Ribosomal DNA Detection.

 

Lee SH, Vigliotti VS, Vigliotti JS, Jones W, Pappu S.

 

300 Seaside Ave, Milford, CT 06460.

 

The DNA of Borrelia burgdorferi spirochetes extracted by ammonium

hydroxide was used as the template for nested polymerase chain

reaction (PCR) amplification of the species-specific 16S

ribosomal DNA (rDNA). The primers were those well known to be

specific for signature sequence amplification of the B

burgdorferi sensu lato 16S ribosomal RNA gene. The positive

293-base-pair nested PCR amplicon was subjected to routine direct

automated Sanger sequencing. A 50-base sequence excised randomly

from the sequencing electrophoretogram between the 2 nested PCR

primer binding sites was sufficient for the Basic Local Alignment

Search Tool

(BLAST) analysis to validate the B burgdorferi sensu lato 16S

rDNA without a reasonable doubt. Nested PCR increased the

sensitivity of DNA detection by 100- to 1,000-fold. DNA sequence

validation based on BLAST algorithms using the GenBank database

practically eliminates any possibility of false-positive results

due to molecular misidentification. This technology may be a

valuable supplement to the current serologic tests for Lyme

disease.

 

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PMID: 20231610 [PubMed - in process]