Gross U, Roggenkamp A, Janitschke K, Heesemann J
Institute of Hygiene and Microbiology, University of Würzburg, Germany.
Eur J Clin Microbiol Infect Dis. 1992 Jan;11(1):33-9. doi: 10.1007/BF01971268.
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.
本研究的目的是提高聚合酶链反应检测生物和临床标本中弓形虫的灵敏度。使用一对引物扩增该寄生虫B1基因的634 bp片段，在含有高浓度伴随宿主细胞的100微升样品悬液中能够检测到10个寄生虫。不同DNA纯化方法的比较表明，用于聚合酶链反应的富含细胞的临床标本在DNA扩增前应用蛋白酶K消化。通过使用所述的样品制备方法和聚合酶链反应，在52例有弓形虫病临床或血清学指征患者的临床标本中，有10例检测到了弓形虫DNA。
