| Literature DB >> 35215221 |
Konstantin Tanida1, Andreas Hahn2, Kirsten Alexandra Eberhardt3,4, Egbert Tannich5,6, Olfert Landt7, Simone Kann8, Torsten Feldt9, Fred Stephen Sarfo10, Veronica Di Cristanziano11, Hagen Frickmann1,2, Ulrike Loderstädt12.
Abstract
In the original publication [...].Entities:
Year: 2022 PMID: 35215221 PMCID: PMC8875394 DOI: 10.3390/pathogens11020256
Source DB: PubMed Journal: Pathogens ISSN: 2076-0817
Details of the real-time PCR assays 1–6, which were included in the test comparison without a reference standard with perfect accuracy for the diagnosis of microsporidia in stool samples. Positive control plasmid inserts are provided in Appendix A.
| PCR 1 | PCR 2 | PCR 3 | PCR 4 | PCR 5 | PCR 6 | |
|---|---|---|---|---|---|---|
| Target specificity | Small subunit ribosomal RNA gene of | Small subunit ribosomal RNA gene of | Small subunit ribosomal RNA gene of | Internal transcribed spacer (ITS) sequence of | Small subunit ribosomal RNA gene of | Internal transcribed spacer (ITS) sequence of the non-target microorganism |
| Amplicon length | 394 base pairs | 280 base pairs | 202 base pairs | 105 base pairs | 227 base pairs | 87 base pairs |
| Cycle number | 50 | 40 | 40 | 50 | 40 | 45 |
| Forward primer 1 | 5′-CACCAGGTTGATTCTGCCTGA-3′ | 5′-CAGGTTGATTCTGCCTGACG-3′ | 5′-CCAGGGTCAAGTCATTCGTT-3′ | 5′-TGTGTAGGCGTGAGAGTGTATCTG-3′ | 5′-CACCAGGTTGATTCTGCCTGAC-3′ | 5′-TCTTGCGCGTTAATGATCCTT-3′ |
| Forward primer 2 | 5′-TCCGGAGAGGGAGCCTGAG-3′ | n.a. | n.a. | n.a. | n.a. | n.a. |
| Reverse primer 1 | 5′-GCTTGCCCTCCAATTGCTTC-3′ | 5′-CCATCTCTCAGGCTCCCTC-3′ | 5′-TATTGTATTGCGCTTGCTGC-3′ | 5′-CATCCAACCATCACGTACCAATC-3′ | 5′-CTAGTTAGGCCATTACCCTAACTACCA-3′ | 5′-AGGTTGCGGGCGGC-3′ |
| Reverse primer 2 | 5′-GACTTGCCCTCCAATCACATG-3′ | n.a. | n.a. | n.a. | n.a. | n.a. |
| Reverse primer 3 | 5′-CCGACTTGCCCTCCAGTAAA-3′ | n.a. | n.a. | n.a. | n.a. | n.a. |
| Reverse primer 4 | 5′-CTTGGCCTCCAATCAATCTCG-3′ | n.a. | n.a. | n.a. | n.a. | n.a. |
| Hybridization probe * | 5′-TGGCAGCAGGCGCGAAACTTGT-3′ | n.a. | 5′-GATGCCCTTAGATATCCTGG-3′ | 5′-CACTGCACCCACATCCCTCACCCTT-3′ | 5′-CTATCACTGAG+C+CGT+CC-3′ | 5′-ACGGAAGAGCTTCGGGGGCCA-3′ |
n.a. = not applicable. * Bases with a plus (+) in front of them are locked nucleic acid (LNA) bases.