In vitro assessment of drug resistance in Plasmodium falciparum in five States of India

Abstract

Background & objectives: In vitro assays are an important tool to assess baseline sensitivity and monitor the drug response of Plasmodium falciparum over time and place and, therefore, can provide background information for the development and evaluation of drug policies. This study was aimed at determining the in vitro sensitivity of P. falciparum isolates to antimalarials.

Methods: The in vitro activity of 108 P. falciparum isolates obtained from five States of India was evaluated using WHO microtest (Mark III) to chloroquine, monodesethylamodiaquine, dihydroartesunate and mefloquine. Samples were collected from the States of Orissa, Jharkhand, Karnataka, Goa and Chhattisgarh from September 2007 to August 2009. In addition, representative samples from different States of India cryopreserved and culture adapted in the Malaria Parasite Bank of National Institute of Malaria Research, New Delhi, were also evaluated.

Results: The proportion of isolates resistant to chloroquine and monodesethylamodiaquine was 44.4 and 25 per cent, respectively. Of the 27 isolates resistant to monodesethylamodiaquine, 16 (59.3%) were cross-resistant to chloroquine. No isolate showed resistance to dihydroartesunate and mefloquine. Isolates from Orissa showed the highest degree of resistance to chloroquine and amodiaquine followed by Jharkhand. Forty two isolates were genotyped for pfcrt T76K chloroquine resistant mutation; mutations were seen in 38 (90.47%) isolates.

Interpretation & conclusions: The Indian P. falciparum isolates showed a high degree of resistance to chloroquine followed by monodesethylamodiaquine. No resistance was recorded to mefloquine and dihydroartesunate.

Fig. 1

Response of a P. falciparum representative isolate to chloroquine.

Fig. 2

Cross-resistance between chloroquine and amodiaquine. CQ, chloroquine ; AQ, amodiaquine; figures succeeding drug name indicate number of isolates.

Fig. 3

Restriction fragment length polymorphism analysis of pfcrt. Representative agarose gel electrophoresis of the pfcrt K76T polymorphism. Presence of the K76T mutation was detected using the ApoI restriction enzyme. ApoI does not cut the mutant allele (lanes 2-17 up & 2-7 down). Lane 1, 100 bp ladder. Lanes 2-17 = Samples M3, M4, M5, M6, M15, M16, M17, M18, M19, M20, M21, M22, M23, M24, M25 Lanes 2-7 = Samples M26, M27, M28, M29 M30, M39.