IMPROVED TESTS FOR BORRELIA - LYME DISEASE

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Int J Med Sci 2013; 10(4):362-376. doi:10.7150/ijms.5698

Research Paper

Improved Culture Conditions for the Growth and Detection of Borrelia from Human Serum

Eva Sapi^1,2 , Namrata Pabbati¹, Akshita Datar¹, Ellen M Davies¹, Amy Rattelle¹, Bruce A Kuo¹

1. Research Division of Advanced Laboratory Services Philadelphia PA, USA;
2. Department of Biology and Environmental Science, University of New Haven, West Haven CT, USA.

How to cite this article:
Sapi E, Pabbati N, Datar A, Davies EM, Rattelle A, Kuo BA. Improved Culture Conditions for the Growth and Detection ofBorrelia from Human Serum. Int J Med Sci 2013; 10(4):362-376. doi:10.7150/ijms.5698. Available fromhttp://www.medsci.org/v10p0362.htm

Abstract

In this report we present a method to cultivate Borrelia spirochetes from human serum samples with high efficiency. This method incorporates improved sample collection, optimization of culture media and use of matrix protein. The method was first optimized utilizing Borrelia laboratory strains, and later by demonstrating growth of Borrelia from sera from fifty seropositive Lyme disease patients followed by another cohort of 72 Lyme disease patients, all of whom satisfied the strict CDC surveillance case definition for Lyme disease. The procedure resulted in positive cultures in 47% at 6 days and 94% at week 16. Negative controls included 48 cases. The positive identification of Borrelia was performed by immunostaining, PCR, and direct DNA sequencing.

In summary, this report provides evidence for the value of a novel method for culturing Borrelia from human serum samples. The inclusion of key components such as modified culture media plus a unique culture environment has resulted in an improvement in the ability to cultivate this organism. This new culture method directly addresses the issue of the low numbers of Borrelia in clinical samples by amplifying their quantity through long term culture in which Borrelia were able to thrive for as long as eight months (data not shown). The versatility of this method allows for samples to be harvested from the culture at any point in time for further study, and it also serves as a source of Borrelia for a variety of direct detection techniques as well as for additional research. Finally, this unique culture method could play an important role in providing useful diagnostic information for select Lyme disease patients who might have tested negatively by other methods.

Acknowledgements

The authors thank Dr. Joseph Burrascano, Jr., Dr. Alan MacDonald and Dr. Parkash Gill for helpful discussion.

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