TABLE 1.

Primers and probes used in the ERA/CRISPR–Cas12 dual system.

Name	Sequence (5′-3′)	Length
Forward Primer 1 (F1)	AAAGAAATCGGACTCGGAGGACAATGGTCAG	31
Forward Primer 2 (F2)*	AAGAAATCGGACTCGGAGGACAATGGTCAG	30
Reverse Primer 1 (R1)	CATAAGGCGCATCGTACAGAATCAGGATCGAG	32
Reverse Primer 2 (R2)*	GTACAGAATCAGGATCGAGGCGGATCATTTGG	32
Forward Primer (F)	GACTCACCGTAGTGGGACACTTCACAAGTACCA	33
Reverse Primer (R)	GTTCGGGTGGGATCATACGTGGTTTGTTGACTG	33
F-Q	/56-FAM/TTATTATT/3BHQ_1/	8
F-B	/56-FAM/TTATTATT/3Biotin/	8

The oligonucleotide sequences of ERA primers and non-specific single-stranded DNA for the trans-cleavage assay used by the ERA/CRISPR–Cas12a dual system for the detection of M. pneumoniae. F and R were the forward and reverse primers designed for gRNA2. The remainder was primers designed for gRNA1. The final sequences are marked with an asterisk (*). F-Q was used in the fluorescence system and F-B in the dipstick system for trans-cleavage detection.