Table 1. List of primers and thermal cycling profiles used in this study.
Target | Primer name | Oligonucleotide sequence | Reference strains for standard curves | PCR profile |
---|---|---|---|---|
All Bacteria | 341f | CCTACGGGAGGCAGCAG | Escherichia coli JCM 1649T | 95°C (5 sec)–60°C (20 sec)–72°C (20 sec)/35 cycles |
534r | ATTACCGCGGCTGCTGG | |||
Actinobacteria | Act920F3 | TACGGCCGCAAGGCTA | Bifidobacterium longum subsp. Longum | 95°C (20 sec)–54°C (20 sec) –72°C (50 sec)/35 cycles |
Act1200R | TCRTCCCCACCTTCCTCCG | JCM 1217T | ||
Bacteroidetes | Bact934F | GGARCATGTGGTTTAATTCGATGAT | Bacteroides fragilis DSM 2151T | 95°C (5 sec)–55°C (30 sec) /35cycles |
Bact1060R | AGCTGACGACAACCATGCAG | |||
Fusobacteia | Fusobacteria F | GATCCAGCAATTCTGTGTGC | Fusobacterium nucleatum subsp. Nucleatum JCM 8532T | 95°C (5 sec)–55°C (20 sec)–72°C (50 sec)/35 cycles |
Fusobacteria R | CGAATTTCACCTCTACACTTGT | |||
Clostridium cluster XIV | CXIV-F1 | GAWGAAGTATYTCGGTATGT | Clostridium clostridioforme JCM 1291T | 95°C (5 sec)–52°C (30 sec)–72°C (30 sec)/35 cycles |
CXIV-R2 | CTACGCWCCCTTTACAC | |||
Bacteroides spp. | HuBac594Bhqf m | GTTGTGAAAGTTTGCGGCTCAACC | Bacteroides fragilis DSM 2151T | 95°C (5 sec)–60°C (30 sec)/35 cycles |
HuBac692r | CTACACCACGAATTCCGCCT | |||
Bifidobacterium spp. | BifiLM26F | GATTCTGGCTCAGGATGAACGC | Bifidobacterium longum subsp. JCM1217T | 95°C (5 sec)–60°C (20 sec)–72°C (20 sec)/35 cycles |
Bif228R | CTGATAGGACGCGACCCCAT | |||
Fusobacterium spp. | * | * | * | 95°C (5 sec)–64°C (30 sec) /35cycles |
Lactobacillus spp. | LactoR’F | CACAATGGACGMAAGTCTGATG | Lactobacillus casei JCM 1134T | 95°C (20 sec)–56°C (20 sec) –72°C (50 sec)/35 cycles |
LBFR | CGCCACTGGTGTTCTTCCAT | |||
Sutterella spp. | ** | ** | ** | 95°C (5 sec)–57°C (20 sec)–72°C (20 sec)/35 cycles |
Clostridium perfringens | Cperf 165F | CGCATAACGTTGAAAGATGG | Clostridium perfringens JCM 1290T | 95°C (5 sec)–60°C (30 sec) /35cycles |
Cperf269R | CCTTGGTAGGCCGTTACCC | |||
Faecalibacterium prausntzii | FPR-2F | GGAGGAAGAAGGTCTTCGG | Faecalibacterium prausnitzii ATCC 27768T | 95°C (5 sec)–57°C (20 sec)–72°C (50 sec)/35 cycles |
Fprau645R | AATTCCGCCTACCTCTGCACT |
*Fusobacterium Detection Kit (TechnoSuruga Laboratory, Shizuoka, Japan). **Sutterella Detection Kit (TechnoSuruga Laboratory).