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[Demonstration of Cryptosporidium parvum in immune suppressed rats using nested PCR]
Hüseyin Can1, Ayşe Caner, Mert Döşkaya
1Ege Üniversitesi Fen Fakültesi, Biyoloji Bölümü, Moleküler Biyoloji Anabilim Dalı, İzmir, Türkiye.
Objective:
In the present study, the aim is to demonstrate Cryptosporidium parvum 18S small-subunit rRNA gene, in lung and stool samples of immune suppressed rats. This gene region is specific for Cryptosporidium spp. and thus can be used in humans for routine diagnostic procedures.
Methods:
Three groups (n=4) of Rattus norvegicus rats were used. The first and second groups were administered dexamethasone, subcutaneously and orally, respectively, for 12 weeks. Rats in the control group were not immune suppressed. Lung and stool specimens were obtained from rats at the end of 12 < sup > th < /sup > week and examined for the presence of C. parvum DNA using Nested PCR.
Results:
C. parvum DNA was demonstrated in lung and stool samples of rats which were immune suppressed by oral dexamethasone. On the other hand, C. parvum DNA was demonstrated only in stool specimens of the rats which were immune suppressed by subcutaneous dexamethasone. No band pattern was observed in the specimens of the control group.
Conclusion:
The results of the study showed that oral dexamethasone administration was more efficient in generating disseminated cryptosporidiosis in rats compared to subcutaneous dexamethasone administration. In addition, Nested PCR targeting 18S small-subunit rRNA gene can be used to detect Cryptosporidium spp. in respiratory and stool specimens of animals and humans.
Insights
Oral dexamethasone effectively induced disseminated cryptosporidiosis in rats, detectable via Nested PCR targeting the Cryptosporidium parvum 18S rRNA gene in lung and stool samples. This method shows promise for diagnosing infections in both animals and humans.
Area of Science:
- Veterinary Parasitology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Cryptosporidium parvum is an opportunistic pathogen causing cryptosporidiosis.
- Immunosuppression can lead to disseminated infections.
- The 18S small-subunit rRNA gene is a reliable target for Cryptosporidium detection.
Purpose of the Study:
- To demonstrate the presence of Cryptosporidium parvum 18S rRNA gene in immunosuppressed rats.
- To evaluate the efficacy of oral versus subcutaneous dexamethasone in inducing disseminated cryptosporidiosis.
- To validate Nested PCR for routine diagnostic applications in humans and animals.
Main Methods:
- Three groups of rats (n=4) were used, with two groups receiving oral or subcutaneous dexamethasone for 12 weeks.
- Lung and stool samples were collected after 12 weeks.
- Nested PCR targeting the 18S small-subunit rRNA gene was employed to detect C. parvum DNA.
Main Results:
- C. parvum DNA was detected in both lung and stool samples of rats treated with oral dexamethasone.
- C. parvum DNA was found only in stool samples of rats treated with subcutaneous dexamethasone.
- No C. parvum DNA was detected in the control group.
Conclusions:
- Oral dexamethasone administration is more effective than subcutaneous administration in establishing disseminated cryptosporidiosis in rats.
- Nested PCR targeting the 18S rRNA gene is a sensitive method for detecting Cryptosporidium spp. in respiratory and fecal specimens.
- This molecular approach is suitable for routine diagnostic procedures in both veterinary and human medicine.
