Genotyping of Plasmodium falciparum gametocytes by reverse transcriptase polymerase chain reaction

被引:47
作者
Menegon, M
Severini, C
Sannella, A
Paglia, MG
Sangaré, D
Abdel-Wahab, A
Abdel-Muhsin, AAM
Babiker, H
Walliker, D
机构
[1] Ist Super Sanita, Lab Parassitol, I-00161 Rome, Italy
[2] IRCCS Lazzaro Spallanzani, Ist Nazl Ric Malattie Infett, Rome, Italy
[3] Fac Med Pharm & Odontostomatol, Malaria Res & Training Ctr, Bamako, Mali
[4] Natl Res Ctr, Res Inst Trop Med, Khartoum, Sudan
[5] Univ Edinburgh, Inst Cell Anim & Populat Biol, Edinburgh, Midlothian, Scotland
[6] Ist Super Sanita, Lab Biol Cellulare, I-00161 Rome, Italy
基金
英国医学研究理事会; 英国惠康基金;
关键词
Plasmodium falciparum; transmission stages; gametocyte carrier; pfg377; mRNA; genetic diversity; RT-PCR assay;
D O I
10.1016/S0166-6851(00)00314-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A molecular assay has been developed for the specific detection and genetic characterisation of Plasmodium falciparum gametocytes in the blood of malaria infected individuals. The assay is based on the reverse transcription and polymerase chain reaction (RT-PCR) amplification of the messenger RNA of gene pfg377, a sexual-stage specific transcript abundantly produced in maturing gametocytes. The gene contains four regions of repetitive sequences, of which region 3 was shown to be the most polymorphic in laboratory clones and held isolates of the parasite. Analysis of samples of malaria infected blood by RT-PCR specific for region 3 has enabled identification of multiple gametocyte-producing clones within single infections. The assay is able to detect gametocytes below the threshold of microscopic detection, and is highly specific for its gametocyte targets also in the presence of a vast excess of asexual forms. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:153 / 161
页数:9
相关论文
共 21 条
[1]   PLASMODIUM-FALCIPARUM - PARASITES DEFECTIVE IN EARLY STAGES OF GAMETOCYTOGENESIS [J].
ALANO, P ;
ROCA, L ;
SMITH, D ;
READ, D ;
CARTER, R ;
DAY, K .
EXPERIMENTAL PARASITOLOGY, 1995, 81 (02) :227-235
[2]   COS cell expression cloning of Pfg377, a Plasmodium falciparum gametocyte antigen associated with osmiophilic bodies [J].
Alano, P ;
Read, D ;
Bruce, M ;
Aikawa, M ;
Kaido, T ;
Tegoshi, T ;
Bhatti, S ;
Smith, DK ;
Luo, C ;
Hansra, S ;
Carter, R ;
Elliott, JF .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1995, 74 (02) :143-156
[3]  
Arez A. P., 1997, Parassitologia (Rome), V39, P65
[4]   Current views on the population structure of Plasmodium falciparum: Implications for control [J].
Babiker, HA ;
Walliker, D .
PARASITOLOGY TODAY, 1997, 13 (07) :262-267
[5]   RANDOM MATING IN A NATURAL-POPULATION OF THE MALARIA PARASITE PLASMODIUM-FALCIPARUM [J].
BABIKER, HA ;
RANFORDCARTWRIGHT, LC ;
CURRIE, D ;
CHARLWOOD, JD ;
BILLINGSLEY, P ;
TEUSCHER, T ;
WALLIKER, D .
PARASITOLOGY, 1994, 109 :413-421
[6]   GENE FLOW AND CROSS-MATING IN PLASMODIUM-FALCIPARUM IN HOUSEHOLDS IN A TANZANIAN VILLAGE [J].
BABIKER, HA ;
CHARLWOOD, JD ;
SMITH, T ;
WALLIKER, D .
PARASITOLOGY, 1995, 111 :433-442
[7]   Detection of low level Plasmodium falciparum gametocytes using reverse transcriptase polymerase chain reaction [J].
Babiker, HA ;
Abdel-Wahab, A ;
Ahmed, S ;
Suleiman, S ;
Ranford-Cartwright, L ;
Carter, R ;
Walliker, D .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1999, 99 (01) :143-148
[8]   Different genetic characteristics of Plasmodium falciparum isolates collected during successive clinical malaria episodes in Senegalese children [J].
Contamin, H ;
Fandeur, T ;
Rogier, C ;
Bonnefoy, S ;
Konate, L ;
Trape, JF ;
MercereauPuijalon, O .
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1996, 54 (06) :632-643
[9]   PFCRK-1, A DEVELOPMENTALLY-REGULATED CDC2-RELATED PROTEIN-KINASE OF PLASMODIUM-FALCIPARUM [J].
DOERIG, C ;
DOERIG, C ;
HORROCKS, P ;
COYLE, J ;
CARLTON, J ;
SULTAN, A ;
ARNOT, D ;
CARTER, R .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1995, 70 (1-2) :167-174
[10]   Shotgun DNA microarrays and stage-specific gene expression in Plasmodium falciparum malaria [J].
Hayward, RE ;
DeRisi, JL ;
Alfadhli, S ;
Kaslow, DC ;
Brown, PO ;
Rathod, PK .
MOLECULAR MICROBIOLOGY, 2000, 35 (01) :6-14