Transactions of the Royal Society of Tropical Medicine and Hygiene (2007) 101, 422—428 available at www.sciencedirect.com journal homepage: www.elsevierhealth.com/journals/trst Evaluation of a real-time PCR assay for malaria diagnosis in patients from Vietnam and in returned travellers Thi Kim Duy Vo a,b , Patricia Bigot c , Pierre Gazin a , Veronique Sinou a,c , Jean Jacques De Pina d , Dinh Chien Huynh b , Francis Fumoux a , Daniel Parzy a,c,* a Universit´ e de la M´ editerran´ ee, EA 864, IFR 48, Marseille, France b University of Hue, 20 Le Loi, Hue, Vietnam c Institut de M´ edecine Tropicale du Service de Sant´ e des Arm´ ees (IMTSSA), Unit´ e de Recherche en Pharmacologie et Physiopathologie Parasitaire (UR3P), Marseille, France d Hˆ opital d’Instruction des Arm´ ees Laveran, Laboratoire de Biologie M´ edicale, Marseille, France Received 22 June 2006; received in revised form 4 August 2006; accepted 13 September 2006 Available online 5 December 2006 KEYWORDS Malaria; Diagnosis; Real-time PCR; Vietnam Summary Real-time PCR diagnosis of malaria has advantages over traditional microscopic methods, especially when parasitaemia is low and when dealing with mixed infections. We have developed a new real-time PCR with specific genes in each Plasmodium species present only in one copy to identify the four pathogenic Plasmodium spp. for humans. The sensitivity was less than 25 parasites/l. No cross-hybridisation was observed with human DNA or among the four Plasmodium spp. Using LightCycler ® PCR and conventional microscopy, we compared the diagnosis of malaria in patients from Vietnam and in returned European travellers with suspicion of malaria. In patients from Vietnam with suspicion of malaria, one mixed infection was observed by PCR only; the remaining data (54 of 55 patients) correlated with microscopy. In 79 patients without symptoms, low parasitaemia was detected in 7 samples by microscopy and in 16 samples by PCR. In returned travellers, PCR results were correlated with microscopy for all four species in 48 of 56 samples. The eight discrepant results were resolved in favour of real-time PCR diagnosis. This new real-time PCR is a rapid, accurate and efficient method for malaria diagnosis in returned travellers as well as for epidemiological studies or antimalarial efficiency trials in the field. © 2006 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved. ∗ Corresponding author. Present address: Centre National de R´ ef´ erence sur le Paludisme Sud, Institut de M´ edecine Tropicale du Service de Sant´ e des Arm´ ees (IMTSSA), UR3P, BP46, 13998 Marseille Arm´ ees, France. Tel.: +33 4 91 15 01 14; fax: +33 4 91 15 01 64. E-mail address: d.parzy@free.fr (D. Parzy). 0035-9203/$ — see front matter © 2006 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.trstmh.2006.09.004