Evaluation of three extraction methods for molecular detection of Schistosoma mansoni infection in human urine and serum samples.
The diagnostic techniques based on polymerase chain reaction (PCR) for the detection of Schistosoma spp. DNA in stool, serum, plasma and urine has shown high sensitivity and specificity solving the problems for the low worm burdens and low transmission rates facing the routine microscopic diagnosis. Since PCR assays require efficient unbiased procedures of extraction and purification of nucleic acids. This study compared the efficiencies of simple, manual and feasible DNA extraction methods; a salting out and resin method, phenol/chloroform method to a commercial extraction kit through PCR analysis of human urine and serum samples spiked with known amounts of adult Schistosoma mansoni DNA confirmed by the application on real samples from patients. In artificially spiked urine gradient, the best mean diagnostic performance was that of salting out and resin then phenol/chloroform and last for the commercial kit. All three methods gave positive results in all tested urine samples which insures comparable high efficiency for DNA detection. In artificially spiked serum gradient, the highest mean diagnostic performance was obtained by the kit then salting out and resin and last by phenol chloroform. In patients' urine samples the phenol/chloroform method showed the highest mean diagnostic performance followed by the resin and then the kit. Using patients' serum samples the resin method showed equal mean diagnostic performance with the phenol/chloroform method which was higher compared to the kit. As regards sensitivity from urine samples the resin and phenol/chloroform showed equal results using artificial gradients and patients' samples. In serum samples the resin and phenol/chloroform showed equal results using artificial gradients while the resin showed better results in patients' samples. It is recommended to extract DNA from urine samples and to use the salting out and resin as a manual DNA extraction method from patients' samples for the molecular diagnosis of Schistosoma mansoni infection.
Department of Parasitology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Rania M
RM
Sarhan
Department of Parasitology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Hanan H
HH
Kamel
Department of Parasitology, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Ghada A
GA
Saad
Internal Medicine Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Ossama A
OA
Ahmed
DNA extraction; PCR; Phenol/chloroform; Salting out resin; Schistosoma mansoni; Serum; Urine