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[Experimental study on detection of Schistosoma japonicum 5D gene by using PCR]
1Department of Infectious Diseases, Hua Shan Hospital, Shanghai Medical University, Shanghai 200040.
Aim:
To search for an effective method to diagnose the schistosomiasis and evaluate the effect of treatment.
Methods:
A PCR protocol was designed to detect the Schistosoma japonicum 5D gene encoding an immunogenic miracidial antigen in cercariae, eggs and adult worms.
Results:
Specific amplification of a 262 bp DNA fragment was achieved by using PCR and a 165 bp DNA fragment by using nest-PCR in cercariae, adult worms and eggs of Schistosoma japonicum. After 40 cycles by PCR, the product could be detected by agarose gel electrophoresis. This PCR protocol was able to detect as few as a single cercariae, a single egg or a pinpoint-sized tissue of adult S. japonicum. Nest-PCR and oligonucleotide probe hybridization could further increase the sensitivity. No specific PCR product was obtained with DNA from human leucocyte, Escherichia coli, Bacillus proteus vulgaris, Streptococcus faecalis and Salmonella typhi.
Conclusion:
The sensitivity and specificity of this PCR protocol are satisfactory.