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Genetic Identification of Intestinal Microsporidia Species in Immunocompromised Patients in Tunisia

Stool samples from 86 immunocompromised patients (51 human immunodeficiency virus (HIV)–infected patients and 35 patients with haematologic malignancies) were systematically screened for intestinal microspordiosis by microscopic examination and polymerase chain reaction (PCR) using universal primer V1/PMP2. Nine samples (10.5%) showed amplification with the predictive size of fragment (6 from HIV–infected patients and 3 from patients with myeloma). Only 5 out of them (all HIV–infected patients) were revealed positive by microscopy. By means of amplicons fragment size, species–specific primers (V1/EB450, V1/IS500) and sequencing, 3 microsporidia species were for the first time identified in Tunisia: Enterocytozoon bieneusi (3 isolates), Encephelitozoon intestinalis (2 isolates), and Encephalitozoon hellem (1 isolate). Systematic use of such sensitive and discriminative molecular tools will contribute to determining the true prevalence of microsporidiosis in Tunisia and to better management of infected immunocompromised subjects.

Received July 4, 2008. Accepted for publication August 20, 2008.

Acknowledgments: The authors thank Saloua Laddab, Essia Ben Hassen, Mohamed Bejaoui, and Tarak Ben Othmane from the Bone Marrow Transplant Center of Tunis; Samah Aissa and Taoufik Ben Chaabene from the Department of Infectious Diseases, and Neyla Ben Romdhane from the Department of Haematology of La Rabta Hospital-Tunis, and all the physicians in charge of the patients for their help in the collection of stools and clinical data. The authors are grateful to Mrs. Rim Essid, Olfa Souissi, Najet Zallagua, and Mr. Adel Rhim from Pasteur Institute of Tunis, and Mr. Sylvestre Billigui from Pitié-Salpétrière hospital, Paris-France, for their technical collaboration.

Financial support: This study was supported by the Ministry of Higher Education, Research and Technology, Tunisia, in the setting of the Research Lab "Parasitoses emergentes" LR 05-SP-03.

^* Address correspondence to Karim Aoun, LR "Parasitoses Émergentes", Laboratoire de Parasitologie, Institut Pasteur de Tunis, 13 Place Pasteur, BP 74, 1002 Tunis Belvedère, Tunisia. E-mail: karim.aoun{at}fmt.rnu.tn

Authors’ addresses: Najla Chabchoub, Aïda Bouratbine, and Karim Aoun, LR "Parasitoses Émergentes", Laboratoire de Parasitologie, Institut Pasteur de Tunis, 13 Place Pasteur, BP 74, 1002 Tunis Belvedère, Tunisia, Tel: 216-71-890-827, Fax: 216-71-791-833. Rim Abdelmalek and Fakher Kanoun, Service des Maladies Infectieuses, la Rabta, 1007 Jebbari Tunis, Tunisia, Tel/Fax: 216-71-578-833. Fethi Mellouli, Service d’Immuno-Hématologie Pédiatrique, Centre de Greffe de Moelle Osseuse, 2 rue Djebel Lakhdhar, Tunis, Tunisia. Marc Thellier, Laboratoire de Parasitologie-Mycologie, Hôpital de la Pitié Salpétrière, 47-83 Boulevard de l’Hôpital, 75 651 Paris, France, Tel: 33-01-42-16-01-84, Fax: 33-1-42-16-01-65.

Reprint requests: Karim Aoun, LR "Parasitoses Émergentes", Laboratoire de Parasitologie, Institut Pasteur de Tunis, 13 Place Pasteur, BP 74, 1002 Tunis Belvedère, Tunisia, Tel: 216-98-699-512, Fax: 216-71-791-833, E-mail: karim.aoun{at}fmt.rnu.tn.