Acta Tropica 93 (2005) 233–237 Combined detection of Brugia malayi and Wuchereria bancrofti using single PCR Kirti Mishra, Dipak Kumar Raj, A.P. Dash 1 , R.K. Hazra ∗ Division of Vector Borne Diseases, Institute of Life Sciences, Nalco Square, Chandrasekharpur, Bhubaneswar 751023, India Received 28 April 2004; received in revised form 22 December 2004; accepted 3 January 2005 Abstract A single step PCR method has been developed for the combined detection of the human filarial parasites, Brugia malayi and Wuchereria bancrofti. Parasites’ DNA were isolated from filaria positive blood samples that were collected from endemic areas. The primers used were Hha1 and Ssp I, which amplified the DNA fragments of 322 bp and 188 bp specific to B. malayi and W. bancrofti, respectively. The sensitivity of the assay was tested with blood and mosquito samples having one W. bancrofti in a pool of 10 B. malayi. The assay was further evaluated on field collected blood and mosquito samples. Use of this assay as a diagnostic tool for the detection of filariasis being the most promising aspect of this study, offers scope for detection of both the parasites even at low levels of infection. © 2005 Elsevier B.V. All rights reserved. Keywords: Polymerase chain reaction; Diagnosis; Brugia malayi; Wuchereria bancrofti Lymphatic filariasis has been the major public health problem in tropical and sub tropical world infecting approximately 120 million people. Of these 90% are caused by Wuchereria bancrofti and 10% are by Brugia malayi. In India, an estimated 454 million people are at risk of the disease (WHO, 2003). With tools currently ∗ Corresponding author. Tel.: +91 674 2300137/2301476; fax: +91 674 2300728. E-mail addresses: apdash2@rediffmail.com (A.P. Dash), rupenangshu@yahoo.co.uk (R.K. Hazra). 1 Director, Malaria Research Center, 22 Shamnath Marg, Delhi 110054, India. available, filariasis in principle could be eliminated (Goodman et al., 2003). But the diagnostic methods, which are currently in use, generally suffer from their own limitations. Microscopic examination, hemofiltra- tion and serodiagnosis that are widely used for diagno- sis purpose, confer little guarantee of species identifi- cation in areas where both the human filarial species co-exist. Diagnosis is an essential element in the management of disease, both at the level of individual patient care and at the level of disease control in populations. In re- cent years considerable effort has been focused on the development of specific diagnostic methods based on 0001-706X/$ – see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.actatropica.2005.01.001