Schistosoma ITS-2 qPCR Detection Protocol

Detection of the Schistosoma-specific internal-transcribed-spacer-2 (ITS-2) target by qPCR was performed at Leiden University Medical Center, as previously described [9 (link), 22 (link)]. Deoxyribonucleic acid extraction of 200 µL CVL, cervical, or vaginal swab fluid was done with QIAamp spin columns (QIAGEN Benelux, Venlo, The Netherlands) according to manufacturer's guidelines. The qPCR output was reported in cycle threshold (Ct) values, and parasite DNA loads were categorized by the following prespecified values: high (Ct < 30), moderate (30 ≤ Ct < 35), low (35 ≤ Ct < 50), and negative (no amplification) [23 (link)].