Molecular Diagnostic Tests for Microsporidia

Table 2

Primers for the identification and sequencing of microsporidian rRNA¹ Genes.

ss^218f^3	CACCAGGTTGATTCTGCC
ss18sf	GTTGATTCTGCCTGACGT
ss350f	CCAAGGA(T/C)GGCAGCAGGCGCGAAA
ss350r	TTTCGCGCCTGCTGCC(G/A)TCCTTG
ss530f	GTGCCAGC(C/A)GCCGCGG
ss530r	CCGCGG(T/G)GCTGGCAC
ss1047r	AACGGCCATGCACCAC
ss1061f	GGTGGTGCATGGCCG
ss1492r	GGTTACCTTGTTACGACTT (Universal primer)
ss1537	TTATGATCCTGCTAATGGTTC
ls212r1	GTT(G/A)GTTTCTTTTCCTC
ls212r2	AATCC(G/A/T/C)(G/A)GTT(G/A)GTTTCTTTTCCTC
ls580r	GGTCCGTGTTTCAAGACGG

1- Primers 18f and 1492r amplify most of the small subunit rRNA of the microsporidia. Primes 530f and 212r1 or 212r2 are used to amplify the small subunit rRNA and the ITS region. The remaining primers are used to sequence, with overlap, the forward and reverse strands of the entire small subunit rRNA and ITS region. ls580r amplifies a variable region of the 5^′ end of the large subunit rRNA gene of many microsporidia (e.g., Nosema and Vairimorpha) but it does not work on all microsporidia. ss1537 allows sequencing closer to the 3^′ end of the small subunit rRNA of many but not all microsporidia. ss350f and ss350r may not be needed for sequencing reactions if 18f and 530r provide sufficient overlap to obtain clear sequence data. 2- ss: denotes primers in the small subunit rRNA gene,  ls: denotes primers in the large subunit rRNA gene, f: forward primer (positive strand),  r: reverse primer (negative strand). 3- Similar to V1 primer. Adapted from [12].