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Open Access Research article

Feasibility of magnetic bead technology for concentration of mycobacteria in sputum prior to fluorescence microscopy

Heidi Albert1*, Patrick J Ademun1, George Lukyamuzi1, Barnabas Nyesiga1, Yukari Manabe2, Moses Joloba34, Stuart Wilson5 and Mark D Perkins6

Author Affiliations

1 FIND, Foundation for Innovative New Diagnostics, Kampala, Uganda

2 Infectious Diseases Institute, Mulago Hospital, Kampala, Uganda

3 Department of Medical Microbiology, Makerere University, Kampala, Uganda

4 National Tuberculosis Reference Laboratory, Wandegaya, Kampala, Uganda

5 Microsens Medtech Ltd., London, UK

6 FIND, Foundation for Innovative New Diagnostics, Geneva, Switzerland

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BMC Infectious Diseases 2011, 11:125  doi:10.1186/1471-2334-11-125

Published: 13 May 2011

Abstract

Background

Direct sputum smear microscopy is the mainstay of TB diagnosis in most low and middle income countries, and is highly specific for Mycobacterium tuberculosis in such settings. However it is limited by low sensitivity, particularly in HIV co-infected patients. Concentration by centrifugation has been reported to be more sensitive than direct smear preparation, but is only suitable for referral laboratories. Simpler concentration methods that could be applied in peripheral laboratories are urgently needed.

Methods

We evaluated the feasibility of an early prototype ligand-coated magnetic bead technology to concentrate M. tuberculosis prior to detection by LED-based fluorescence microscopy compared with direct Ziehl-Neelsen microscopy and direct and concentrated fluorescence microscopy in a reference laboratory in Kampala, Uganda. Results were compared with MGIT 960 liquid culture and Lowenstein-Jensen culture.

Results

Compared to culture, concentrated FM had significantly higher sensitivity than direct ZN (74.8% and 51.4%), magnetic bead-FM (65.4%) and direct FM (58.9%). The sensitivity of magnetic bead FM was significantly higher than direct ZN (p < 0.001) but not significantly higher than direct FM (p = 0.210). The specificity of magnetic bead FM and concentrated FM was significantly lower than direct ZN (88.6%, 94.3% and 98.9% respectively) and direct FM (99.4%). There was no significant difference in specificity between magnetic bead FM and concentrated FM. Allowing for blinded resolution of discrepant results, the specificity of magnetic bead FM increased to 93.1%. Direct microscopy was simpler than concentrated FM and Magnetic bead FM which both had a similar number of steps.

Conclusion

The sensitivity of the early prototype magnetic bead FM was lower than concentrated FM, similar to direct FM, and significantly higher than direct ZN. Both magnetic bead and concentration by centrifugation led to reduced specificity compared with the direct smear methods. Some magnetic bead FM false positive results were not easily explained and should be further investigated. The prototype version of the magnetic bead procedure tested here was of similar complexity to concentration by centrifugation. As such, if the sensitivity of the magnetic bead FM could be improved in future versions of the technology, this may offer a viable alternative to centrifugation.