Staff Members - Benha University
Assessment of diagnostic accuracy of a commercial Direct Blood PCR kit for detection of Schistosoma mansoni infection in mice compared with pre extracted PCR assay.
• 2016
Publication Information
Authors
Eraky Maysa A. and Aly Nagwa S. M.
Keywords
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Journal
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Publisher
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Volume
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Issue
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Pages
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publication.type
Local
Paper Link
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Supplementary Materials
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Abstract
Background
Different diagnostic techniques have been used in the diagnosis of schistosomiasis. However, they are far from ideal regarding its early diagnosis. PCR techniques have been tried to improve the direct detection of schistosomiasis.
Objective
The aim of the present study was to evaluate the diagnostic performance of direct amplification of Schistosoma mansoni DNA in the early prepatent period in experimentally infected mice by PCR technique using unextracted DNA as PCR template compared with pre-extracted S. mansoni DNA samples.
Materials and methods
Mice were infected by 100±10 S. mansoni cercariae. Three mice were sacrificed every 3 or 4 days for 5 weeks. Whole blood samples were collected for direct amplification without prior extraction. Serum samples were pooled, and the extracted DNA was detected by using the KAPA blood PCR kit and conventional PCR methods. The diagnostic performance was compared between the two methods.
Results
The results showed that the diagnosis of S. mansoni utilizing pre-extracted DNA was superior to direct amplification of DNA, bypassing nucleic acid extraction which failed to detect S. mansoni DNA in any of the examined samples. Pre-extracted DNA was detected in all samples from the second day after infection by using the two PCR techniques.
Conclusion
These results indicate that S. mansoni infection cannot be efficiently detected directly by using the PCR technique without pre-extraction of DNA from whole blood samples using the KAPA blood PCR kit.
Different diagnostic techniques have been used in the diagnosis of schistosomiasis. However, they are far from ideal regarding its early diagnosis. PCR techniques have been tried to improve the direct detection of schistosomiasis.
Objective
The aim of the present study was to evaluate the diagnostic performance of direct amplification of Schistosoma mansoni DNA in the early prepatent period in experimentally infected mice by PCR technique using unextracted DNA as PCR template compared with pre-extracted S. mansoni DNA samples.
Materials and methods
Mice were infected by 100±10 S. mansoni cercariae. Three mice were sacrificed every 3 or 4 days for 5 weeks. Whole blood samples were collected for direct amplification without prior extraction. Serum samples were pooled, and the extracted DNA was detected by using the KAPA blood PCR kit and conventional PCR methods. The diagnostic performance was compared between the two methods.
Results
The results showed that the diagnosis of S. mansoni utilizing pre-extracted DNA was superior to direct amplification of DNA, bypassing nucleic acid extraction which failed to detect S. mansoni DNA in any of the examined samples. Pre-extracted DNA was detected in all samples from the second day after infection by using the two PCR techniques.
Conclusion
These results indicate that S. mansoni infection cannot be efficiently detected directly by using the PCR technique without pre-extraction of DNA from whole blood samples using the KAPA blood PCR kit.