TABLE 1.
Evaluation of nested PCR amplification and restriction enzyme digestion of a segment within the 18S rRNA of Cryptosporidium for species differentiationa
| Test organism (strain, genotype) | PCR product (sizeb[bp]) obtained from:
|
Results of RFLP digestion with:
|
||
|---|---|---|---|---|
| External primers | Nested primers | VspI | DraII | |
| C. parvum (Peru, 1) | + (844) | + (593) | + | − |
| C. parvum (Iowa, 2) | + (840) | + (590) | − | − |
| C. parvum (KSU-1, 2) | + (840) | + (590) | − | ND |
| C. serpentis (KSU-2) | + (836) | + (583) | − | + |
| C. andersoni (KSU-3) | − | − | ND | ND |
| C. parvum (KSU-4, 2) | + (840) | + (590) | − | ND |
| C. baileyi | + (831) | + (579) | − | + |
| C. muris (108735) | − | − | ND | ND |
| Cyclospora cayetanensis | + (883) | − | ND | ND |
| Eimeria neischulzi | + (885) | − | ND | ND |
| Encephalitozoon intestinalis | − | − | ND | ND |
| Giardia duodenalis | − | − | ND | ND |
| Neospora caninum | +c | − | ND | ND |
| Bacillus subtilis | − | − | ND | ND |
| Escherichia coli | − | − | ND | ND |
a
+, amplification or digestion; −, no amplification or digestion; ND, not done.
b
Amplicon size based on the following representative GenBank accession numbers: C. parvum genotypes 1 and 2, AF093491 and AF164102, respectively; C. baileyi, L19068; C. serpentis, AF151376; C. cayetanensis, AF111183; E. neischulzi, U40263.
c
Multiple PCR fragments observed.