Abstract
Diagnosis of Schistosoma haematobium relies primarily on microscopical analysis of urine. The method is time consuming and requires some expertise. Genus-specific real-time PCRs have been developed, but we still observed low sensitivity. In the present study, in order to achieve a more sensitive DNA detection of eggs of S. haematobium in urine samples, we wanted to develop a novel protocol of DNA extraction using mechanic disruption of eggs by bead beating as supplementary step. We tested Schistosoma spp. internal transcribed spacer 2 real-time PCR after both methods with and without bead beating. First, we preliminary assessed the DNA detection after bead beating using dilution of 2, 10, 50, and 90 eggs/10 mL, and the Ct value analysis showed significant improved DNA detection per each point of egg concentration using the novel supplementary step. Twenty microscopy positive and five microscopy negative urine samples were used to validate the procedure. All urines came from imported cases and admitted at center for tropical medicine, and were examined by microscopy. PCR results after novel method with bead beating showed 100% to be positive for S. haematobium, compared with 85% positive by our standard extraction procedure. Results confirmed mechanic disruption of eggs by bead beating before DNA extraction to be highly effective method for the detection of S. haematobium DNA in urine.
| Original language | English |
|---|---|
| Pages (from-to) | 683-686 |
| Number of pages | 4 |
| Journal | Parasitology Research |
| Volume | 118 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 14 2019 |
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Keywords
- Bead beating
- DNA
- Real-time PCR
- Schistosoma haematobium
ASJC Scopus subject areas
- Parasitology
- veterinary(all)
- Insect Science
- Infectious Diseases
Cite this
Improved detection of DNA Schistosoma haematobium from eggs extracted by bead beating in urine. / Pomari, Elena; Perandin, Francesca; La Marca, Giulia; Bisoffi, Zeno.
In: Parasitology Research, Vol. 118, No. 2, 14.02.2019, p. 683-686.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Improved detection of DNA Schistosoma haematobium from eggs extracted by bead beating in urine
AU - Pomari, Elena
AU - Perandin, Francesca
AU - La Marca, Giulia
AU - Bisoffi, Zeno
PY - 2019/2/14
Y1 - 2019/2/14
N2 - Diagnosis of Schistosoma haematobium relies primarily on microscopical analysis of urine. The method is time consuming and requires some expertise. Genus-specific real-time PCRs have been developed, but we still observed low sensitivity. In the present study, in order to achieve a more sensitive DNA detection of eggs of S. haematobium in urine samples, we wanted to develop a novel protocol of DNA extraction using mechanic disruption of eggs by bead beating as supplementary step. We tested Schistosoma spp. internal transcribed spacer 2 real-time PCR after both methods with and without bead beating. First, we preliminary assessed the DNA detection after bead beating using dilution of 2, 10, 50, and 90 eggs/10 mL, and the Ct value analysis showed significant improved DNA detection per each point of egg concentration using the novel supplementary step. Twenty microscopy positive and five microscopy negative urine samples were used to validate the procedure. All urines came from imported cases and admitted at center for tropical medicine, and were examined by microscopy. PCR results after novel method with bead beating showed 100% to be positive for S. haematobium, compared with 85% positive by our standard extraction procedure. Results confirmed mechanic disruption of eggs by bead beating before DNA extraction to be highly effective method for the detection of S. haematobium DNA in urine.
AB - Diagnosis of Schistosoma haematobium relies primarily on microscopical analysis of urine. The method is time consuming and requires some expertise. Genus-specific real-time PCRs have been developed, but we still observed low sensitivity. In the present study, in order to achieve a more sensitive DNA detection of eggs of S. haematobium in urine samples, we wanted to develop a novel protocol of DNA extraction using mechanic disruption of eggs by bead beating as supplementary step. We tested Schistosoma spp. internal transcribed spacer 2 real-time PCR after both methods with and without bead beating. First, we preliminary assessed the DNA detection after bead beating using dilution of 2, 10, 50, and 90 eggs/10 mL, and the Ct value analysis showed significant improved DNA detection per each point of egg concentration using the novel supplementary step. Twenty microscopy positive and five microscopy negative urine samples were used to validate the procedure. All urines came from imported cases and admitted at center for tropical medicine, and were examined by microscopy. PCR results after novel method with bead beating showed 100% to be positive for S. haematobium, compared with 85% positive by our standard extraction procedure. Results confirmed mechanic disruption of eggs by bead beating before DNA extraction to be highly effective method for the detection of S. haematobium DNA in urine.
KW - Bead beating
KW - DNA
KW - Real-time PCR
KW - Schistosoma haematobium
UR - http://www.scopus.com/inward/record.url?scp=85056320710&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85056320710&partnerID=8YFLogxK
U2 - 10.1007/s00436-018-6137-7
DO - 10.1007/s00436-018-6137-7
M3 - Article
C2 - 30417247
AN - SCOPUS:85056320710
VL - 118
SP - 683
EP - 686
JO - Zeitschrift fur Parasitenkunde
JF - Zeitschrift fur Parasitenkunde
SN - 0044-3255
IS - 2
ER -