[Tag primer-nested/ multiplex PCR for detection of Plasmodium falciparum and Plasmodium vivax].
OBJECTIVE
To establish a sensitive, simple to use and low noise nested/multiplex PCR for simultaneously detection of Plasmodium falciparum (Pf) and Plasmodium vivax (P.v).


METHODS
The tag primer amplification technique, software Primer Premier 5.0, NCBI-BLAST web resources and the matrix test were used to optimize the nested/multiplex PCR for detection of P.f and P.v with filter paper blood samples taken from malaria patients diagnosed by microscopy, and the results of the optimized nested/multiplex PCR and microscopy were evaluated.


RESULTS
The sensitivity of the optimized PCR, determined by the examination of imitative filter paper blood samples, was about 1-2 parasites / microl for P.f and 5-10 parasites / microl for P.v. Primer-dimer and other PCR noise were removed. When 71 field filter paper blood samples taken from microscopically diagnosed patients (24 P.f, 47 P.v) were examined, the concordance between the optimized PCR and microscopy was 875% for Pf and 100% for P.v.


CONCLUSIONS
The nested/multiplex PCR optimized by tag primer amplification technique is simple, with low noise and being able to detect Pf and P.v simultaneously. It is more sensitive in detecting cases with low parasitaemia and more accurate in identifying Plasmodium species than microscopy.
Huang
Tian-Yi
TY
Guizhou Provincial Center for Diseases Control and Prevention, Guiyang 550004, China.
Wang
Shi-Hai
SH
Li
Xue-Ming
XM
Guo
Chuan-Kun
CK
Xu
Jian-Jun
JJ
Tang
Li-Na
LN
Lu
Li-Dan
LD
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000097:blood; D006801:Humans; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D016054:DNA, Protozoan