A two-tiered serological test based on an enzyme-linked immunosorbent assay (ELISA) or an indirect immunofluorescence assay (IFA) followed by a western blot of samples that test positive or equivocal by ELISA or IFA, or alternatively the culture confirmation of BB, are presently recommended by the US Centers for Disease Control (CDC) for laboratory diagnosis. The laboratory diagnosis of Lyme disease is also difficult and the appropriate procedure is a matter of current debate. Patients with chronic fatigue syndrome, fibromyalgia, immune disorders, multiple sclerosis or infections with other microorganisms may present with Lyme disease-like symptoms, adding to the importance of accurately diagnosing Lyme disease in the laboratory. Cardiac, dermatologic, musculoskeletal and neurological manifestations may be evident in advanced stages of Lyme disease. Other common symptoms of acute BB infections are arthralgia, fatigue, fever, headache, mild stiff neck and myalgia. However EM is observed in only 60–80% of BB infections. A ‘bulls-eye’ rash or erythema migrans (EM) is generally the best indicator of an early acute infection. Lyme disease caused by the spirochete bacterium Borrelia burgdorferi (BB) is endemic in many parts of the world and the most prevalent tick-borne disease in Europe and the United States. Use of the LM-PCR assay and optimization of current CDC serological criteria can improve the diagnosis of Lyme disease. 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. 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. The LM-PCR assay was highly specific for B. 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. burgdorferi DNA followed by PCR amplification of flagellin and OspA gene fragments and their detection by southern dot blots. The LM-PCR assay utilizes specific DNA hybridization to purify B. 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 detecting B. PCR tests have previously shown poor sensitivity and are not routinely used for diagnosis. burgdorferi can be weak or variable, or alternatively, the slow and inefficient culture confirmation of B. Laboratory diagnosis of Lyme disease is difficult and presently dependent on detecting Borrelia burgdorferi-specific antibodies in patient serum with the disadvantage that the immune response to B.

Dot blot results