Development of a sensitive PCR-dot blot assay to supplement serological tests for diagnosing Lyme disease

AbstractLaboratory diagnosis of Lyme disease is difficult and presently dependent on detectingBorrelia burgdorferi-specific antibodies in patient serum with the disadvantage that the immune response toB. burgdorferi can be weak or variable, or alternatively, the slow and inefficient culture confirmation ofB. burgdorferi. PCR tests have previously shown poor sensitivity and are not routinely used for diagnosis. We developed a sensitive and specific Lyme Multiplex PCR-dot blot assay (LM-PCR assay) applicable to blood and urine samples to supplement western blot (WB) serological tests for detectingB. burgdorferi infection. The LM-PCR assay utilizes specific DNA hybridization to purifyB. burgdorferi DNA followed by PCR amplification offlagellin andOspA gene fragments and their detection by southern dot blots. Results of the assay on 107 and 402 clinical samples from patients with suspected Lyme disease from Houston, Texas or received at the IGeneX laboratory in Palo Alto, California, respectively, were analyzed together with WB findings. The LM-PCR assay was highly specific forB. burgdorferi. In the Texas samples, 23 (21.5%) patients antibody-negative in WB assays by current US Centers for Disease Control (CDC) recommended criteria were positive by LM-PCR performed on urine, serum or whole blood samples. With IGeneX samples, of the 402 LM-PCR positive blood samples, only 70 met the CDC criteria for positive WBs, while 236 met IGeneX criteria for positive WB. Use of the LM-PCR ass...
Source: European Journal of Clinical Microbiology and Infectious Diseases - Category: Microbiology Source Type: research