Multiplex PCR and oligonucleotide microarray for detection of single-nucleotide polymorphisms associated with Plasmodium falciparum drug resistance.
Drug resistance in Plasmodium falciparum is a serious public health threat in the countries where this organism is endemic. Since resistance has been associated with specific single-nucleotide polymorphisms (SNPs) in parasite genes, molecular markers are becoming useful surrogates for monitoring the emergence and dispersion of drug resistance. In this study, a multiplex PCR (mPCR) and oligonucleotide microarray method was developed for the detection of these SNPs in genes encoding chloroquine resistance transporter (Pfcrt), multidrug resistance 1 (Pfmdr1), dihydrofolate reductase (Pfdhfr), dihydropteroate synthetase (Pfdhps), and ATPase 6 (PfATPase6) of P. falciparum. The results show that DNA microarray technology, combined with mPCR, is a promising and time-saving tool that supports conventional detection methods, allowing sensitive, accurate, simultaneous analysis of the SNPs associated with drug resistance in P. falciparum.
Zhang
Guo Qing
GQ
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Center for Malaria, Schistosomiasis, and Filariasis, 207 Rui Jin Er Road, Shanghai 200025, China.
Guan
Ya Yi
YY
Sheng
Hai Hui
HH
Zheng
Bin
B
Wu
Song
S
Xiao
Hua Sheng
HS
Tang
Lin Hua
LH
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics*; D004094:Dihydropteroate Synthase / Q000235:genetics; D004351:Drug Resistance; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics; D046228:Microarray Analysis; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D015800:Protozoan Proteins / Q000235:genetics
D000252:Calcium-Transporting ATPases; D017931:DNA Primers; D016054:DNA, Protozoan; D004094:Dihydropteroate Synthase; D026901:Membrane Transport Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D015800:Protozoan Proteins; C513500:ATP6 protein, Plasmodium falciparum; C513498:Mdr1 protein, Plasmodium falciparum; C416369:PfCRT protein, Plasmodium falciparum