Two-Year Evaluation of Borrelia burgdorferi Culture and Supplemental Tests for Definitive Diagnosis of Lyme Disease

Abstract

Lyme disease is usually diagnosed and treated based on clinical manifestations. However, laboratory testing is useful for patients with confusing presentations and for validation of disease in clinical studies. Although cultivation of Borrelia burgdorferi is definitive, prior investigations have shown that no single test is optimal for Lyme disease diagnosis. We applied high-volume blood culture, skin biopsy culture, PCR, and serodiagnosis to a cohort of patients with suspected Lyme disease acquired in Maryland and southern Pennsylvania. The study was performed to confirm the relative utility of culture and to identify laboratory testing algorithms that will supplement clinical diagnosis. Overall, 30 of 86 patients (35%) were culture positive, whereas an additional 15 of 84 (18%) were seropositive only (51% total sero-and culture positive), and PCR on skin biopsy identified 4 additional patients who were neither culture nor seropositive. Among 49 laboratory test-positive patients, the highest sensitivity (100%) for diagnosis was obtained when culture, skin PCR, and serologic tests were used, although serologic testing with skin PCR was almost as sensitive (92%). Plasma PCR was infrequently positive and provided no additional diagnostic value. Although culture is definitive and has a relatively high sensitivity, the results required a mean of 3.5 weeks to recovery. The combination of acute-phase serology and skin PCR was 75% sensitive, offering a practical and relatively rapid alternative for confirming clinical impression. The full battery of tests could be useful for patients with confusing clinical signs or for providing strong laboratory support for clinical studies of Lyme disease.

Diagnostic challenges of early Lyme disease: Lessons from a community case series

“…No confirmatory tests (such as PCR or culture) are available to community practitioners despite their potential utility in early diagnosis, and serologies offer a 60% false negative rate in the first weeks of infection [4,24,28]. Accurate diagnosis of EM requires an experienced clinical impression of the rash's appearance [5].…”

“…Clinical diagnosis of EM was made by a consulting physician (JA) with a high rate of specificity in EM diagnosis [24]. …”

The Pathogenesis of Lyme Neuroborreliosis: From Infection to Inflammation

“…also uses this path. In accordance with this, borrelia can be cultivated in up to 35% to 45% of plasma samples from patients with early Lyme disease in the US (70,71). It has to be kept in mind that the effective prevalence of borrelia in the blood of patients will be even higher, as the sensitivity of culture methods can never achieve 100%.…”

Development of a sensitive molecular diagnostic assay for detectingBorrelia burgdorferiDNA from blood of Lyme disease patients by digital PCR

“…Inclusion of various steps to improve sensitivity: pre-analytical processing, pre-amplification of Borreliaspecific gene targets prior to PCR and using the dPCR platform has led to the development of a robust assay to detect B. burgdorferi DNA from the blood of patients. By employing these methods, we were able to overcome the loss of assay sensitivity due to the presence of very low number of spirochetes in the blood stream of patients [11,13,22,23]. The results from this study indicate that our dPCR assay has succeeded where previous assays involving PCR technology had failed to achieve the required sensitivity to become a clinically approved diagnostic assay for LD [8,9].…”