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Journal of Clinical Microbiology, November 2006, p. 3887-3893, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.00969-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Detection of Brugia Parasite DNA in Human Blood by Real-Time PCR{triangledown}

Ramakrishna U. Rao,1 Gary J. Weil,1 Kerstin Fischer,1 Taniawati Supali,2 and Peter Fischer1*

Department of Internal Medicine, Infectious Diseases Division, Washington University School of Medicine, St. Louis, Missouri,1 Department of Parasitology, Faculty of Medicine, University of Indonesia, Jakarta, Indonesia2

Received 9 May 2006/ Returned for modification 6 July 2006/ Accepted 28 August 2006

Brugian filariasis (caused by the nematodes Brugia malayi and B. timori) is an important cause of disability in Southeast Asia. Improved diagnostic tests are needed for filariasis elimination programs (to identify areas of endemicity and to monitor progress) and for diagnosis of the disease in infected individuals. We have developed and evaluated two real-time PCR assays for detecting Brugia DNA in human blood and compared the results of these assays to those of "gold standard" assays. One assay uses a TaqMan probe (TaqM) to amplifiy a 320-bp "HhaI repeat" DNA sequence. The other assay uses a minor groove binding probe (MGB) and modified nucleotides in primers (Eclipse MGB) to amplify a 120-bp fragment of the HhaI repeat. This assay detects 22 copies of the target sequence, and it is more sensitive than the TaqM assay. Both assays were evaluated with human blood samples from two different areas of endemicity. The MGB assay was as sensitive as membrane filtration and microscopy for the detection of B. malayi infection in 57 blood samples recovered at night from patients in Sulawesi, Indonesia. The MGB assay also detected parasite DNA in 17 of 31 (55%) of microfilaria-negative day blood samples from these subjects. This test was more sensitive than the conventional and the TaqM PCRs (and was almost as sensitive as night blood membrane filtration) for the detection of infection in 52 blood samples recovered at night from individuals in an area of B. timori endemicity on Alor Island, Indonesia, where microfilaria-positive individuals had low densities after mass treatment. Thus, the Eclipse MGB real-time PCR assay is a sensitive means of detecting Brugia parasite DNA in human blood.

* Corresponding author. Mailing address: Department of Internal Medicine, Infectious Diseases Division, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8051, St. Louis, MO 63110. Phone: (314) 454-7876. Fax: (314) 454-5293. E-mail: Pufische{at}im.wustl.edu.

{triangledown} Published ahead of print on 6 September 2006.

Journal of Clinical Microbiology, November 2006, p. 3887-3893, Vol. 44, No. 11
0095-1137/06/$08.00+0     doi:10.1128/JCM.00969-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Thanchomnang, T., Intapan, P. M., Lulitanond, V., Choochote, W., Manjai, A., Prasongdee, T. K., Maleewong, W. (2008). Rapid Detection of Brugia malayi in Mosquito Vectors Using a Real-time Fluorescence Resonance Energy Transfer PCR and Melting Curve Analysis. Am J Trop Med Hyg 78: 509-513 [Abstract] [Full Text]  
  • FISCHER, P., ERICKSON, S. M., FISCHER, K., FUCHS, J. F., RAO, R. U., CHRISTENSEN, B. M., WEIL, G. J. (2007). PERSISTENCE OF BRUGIA MALAYI DNA IN VECTOR AND NON-VECTOR MOSQUITOES: IMPLICATIONS FOR XENOMONITORING AND TRANSMISSION MONITORING OF LYMPHATIC FILARIASIS. Am J Trop Med Hyg 76: 502-507 [Abstract] [Full Text]  


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