Improved sensitivity of the polymerase chain reaction for detection of Toxoplasma gondii in biological and human clinical specimens.
The aim of the present study was to improve the sensitivity of the polymerase chain reaction for detection of Toxoplasma gondii in biological and clinical specimens. Using a pair of primers amplifying a 634 bp fragment of the B1 gene of this parasite, it was possible to detect ten parasites in 100 microliters of sample suspensions containing a high concentration of concomitant host cells. A comparison of different DNA purification methods indicated that cell-rich clinical specimens intended for use as samples for the polymerase chain reaction should be digested with proteinase K prior to DNA amplification. By using the described sample preparation methods and the polymerase chain reaction, Toxoplasma gondii DNA was demonstrated in ten of 52 clinical specimens of patients with clinical or serological indications of toxoplasmosis.
Gross
U
U
Institute of Hygiene and Microbiology, University of Würzburg, Germany.
Roggenkamp
A
A
Janitschke
K
K
Heesemann
J
J
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis / Q000378:metabolism; D019286:Endopeptidase K; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000175:diagnosis*
D016054:DNA, Protozoan; D009838:Oligodeoxyribonucleotides; D012697:Serine Endopeptidases; D019286:Endopeptidase K