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Journal of Clinical Microbiology, October 1999, p. 3260-3264, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Usefulness of Seminested Multiplex PCR in Surveillance of Imported Malaria in Spain

J. M. Rubio,1 A. Benito,1,* P. J. Berzosa,1 J. Roche,1 S. Puente,2 M. Subirats,2 R. López-Vélez,3 L. García,1 and J. Alvar1,dagger

Unidad de Investigación en Medicina Tropical y Salud Internacional, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda,1 and Department of Tropical Medicine and Infectious Diseases, Hospital Carlos III, Instituto de Salud Carlos III,2 and Unit of Tropical Medicine, Hospital Ramon y Cajal,3 Madrid, Spain

Received 29 April 1999/Returned for modification 17 June 1999/Accepted 19 July 1999

The use of a new PCR-based method for the diagnosis of malaria in the Spanish Malaria Reference Laboratory has promoted an increase in confirmed cases of malaria. From August 1997 to July 1998, a total of 192 whole-blood samples and 71 serum samples from 168 patients were received from the hospitals of the Spanish National Health System. Most of the patients came from west-central African countries (85%). This molecular method showed more sensitivity and specificity than microscopy, detecting 12.4% more positive samples than microscopy and 13% of mixed infections undetectable by Giemsa stain. Plasmodium falciparum was the main species detected, with 68% of the total positive malaria cases, followed by Plasmodium malariae (29%), Plasmodium vivax (14%), and Plasmodium ovale (7%), including mixed infections in all cases. This report consists of the first wide, centralized survey of malaria surveillance in Spain. The reference laboratory conducted the analysis of all imported cases in order to detect trends in acquisition. The use of a seminested multiplex PCR permitted confirmation of the origins of the infections and the Plasmodium species involved and confirmation of the effectiveness of drug treatments. This PCR also allowed the detection of the presence in Spain of primaquine-tolerant P. vivax strains from west-central Africa, as well as the detection of a P. falciparum infection induced by transfusion.

* Corresponding author. Mailing address: Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo km 2, Majadahonda, 28220 Madrid, Spain. Phone: 34-91-5097901. Fax: 34-91-5097966. E-mail: abenito{at}isciii.es.

dagger Present address: Christ's College, Cambridge CB2 3BU, United Kingdom.

Journal of Clinical Microbiology, October 1999, p. 3260-3264, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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