Volume 18, Number 8—August 2012
Letter
Identification of Cause of Posttransplant Cachexia by PCR
Joelle Guitard
, Sophie Edouard, Hubert Lepidi, Christine Segonds, Marion Grare, Marie-Laure Ranty-Quintyn, Isabelle Rouquette, Olivier Cointault, Lionel Rostaing, Nassim Kamar, and Florence Fenollar
, Sophie Edouard, Hubert Lepidi, Christine Segonds, Marion Grare, Marie-Laure Ranty-Quintyn, Isabelle Rouquette, Olivier Cointault, Lionel Rostaing, Nassim Kamar, and Florence FenollarTable A1
Approach used to determine the cause of posttransplant cachexia in a patient*
| Pathogen | Sequence target | Primers, 5′ → 3′ | Probes/identification |
|---|---|---|---|
| Molecular tool to detect and identify Tropheryma whipplei | |||
| Real-time PCR | |||
| T. whipplei | Repeated sequence | Twhi2F: TGAGGATGTATCTGTGTATGGGACA | 6-FAM-GAGAGATGGGGTGCAGGACAGGG-TAMRA |
| Twhi2R: TCCTGTTACAAGCAGTACAAAACAAA | |||
| T. whipplei | Repeated sequence | Twhi3F: TTGTGTATTTGGTATTAGATGAAACAG | 6-FAM-GGGATAGAGCAGGAGGTGTCTGTCTGG-TAMRA |
| Twhi3R: CCCTACAATATGAAACAGCCTTTG | |||
| Molecular tools to detect and identify Mycobacterium spp. | |||
| Step 1: Real-time PCR | |||
| Mycobacterium spp. | ITS | ITSd: GGGTGGGGTGTGGTGTTTGA ITSr: CAAGGCATCCACCATGCGC | 6-FAM-TGGATAGTGGTTGCGAGCATC-TAMRA |
| M. tuberculosis | ITS | ITSd: GGGTGGGGTGTGGTGTTTGA ITSr: CAAGGCATCCACCATGCGC | 6-FAM-GCTAGCCGGCAGCGTATCCAT-TAMRA |
| M. avium | ITS | ITSd: GGGTGGGGTGTGGTGTTTGA ITSr: CAAGGCATCCACCATGCGC | 6-FAM-GGCCGGCGTTCATCGAAAT-Mgb |
| Step 2: Classical PCR | |||
| Mycobacterium spp. | rpoB | MycoF: GGCAAGGTCACCCCGAAGGG MycoR: AGCGGCTGCTGGGTGATCATC | Sequencing |
| Housekeeping gene | β-actin | ActinF: CATGCCATCCTGCATCTGGA ActinR: CCGTGGCCATCTCTTGCTCG | 6-FAM-CGGGAAATCGTGCGTGACATTAAG-TAMRA |
*ITS, internal transcribed spacer; rpoB, RNA polymerase B.
