TABLE 7.

Primer pairs and hybridization probes used for diagnosis and species differentiation of human microsporidia

Primer pair sequence	Primer designation	Hybridization probe
5′-CACCAGGTTGATTCTGCCTGAC-3′	V1	5′-TGTTGCGGTAATTTGGTCTCTGTGTGTAAA-3′
5′-ACTCAGGTGTTATACTCACGTC-3′	EB450
5′-GCCTGACGTAGATGCTAGTC-3′		5′-GTTCAATAGCGATGAGTTTGCTAATGTT-3′
5′-ATGGTTCTCCAACTGAAACC-3′
5′-GAAACTTGTCCACTCCTTACG-3′	EBIEF1
5′-CCATGCACCACTCCTGCCATT-3′	EBIER1
5′-CACCAGGTTGATTCTGCCTGAC-3′	V1
5′-CAGCATCCACCATAGACAC-3′	Mic3
5′-TCAGTTTTGGGTGTGGTATCGG-3′	Eb.gc
5′-GCTACCCATACACACATCATTC-3′	Eb.gt
5′-CACCAGGTTGATTCTGCCTGAC-3′	V1	5′-TGTTGATGAACCTTGTGG-3′
5′-CTCGCTCCTTTACACTCGAA-3′	SI500	5′-TGGTAGTTTAGGGTAATGGCCTAACTAGGCG-3′
5′-GGGGGCTAGGAGTGTTTTTG-3′		5′-TTTGGGGGATTATGTCCTGATGTGGAT-3′
5′-CAGCAGGCTCCCTCGCCATC-3′
5′-TTTCGAGTGTAAAGGAGTCGA-3′	SINTF1
5′-CCGTCCTCGTTCTCCTGC-3′	SINTR
5′-CACCAGGTTGATTCTGCCTGAC-3′	V1
5′-CCTGCCCGCTTCAGAACC-3′	Sep1
5′-TGAGAAGTAAGATGTTTAGCA-3′	EHEL-F
5′-GTAAAAACACTCTCACACTCA-3′	EHEL-R
5′-ATGAGAAGTGATGTGTGTGTGCG-3′	ECUN-F
5′-TGCCATGCACTCACAGGCATC-3′	ECUN-R
5′-CACCAGGTTGATTCTGCCTGAC-3′	PMP1 (V1)
5′-CCTCTCCGGAACCAAACCTG-3′	PMP2
5′-CACCAGGTTGATTCTGCC-3′	C1
5′-GTGACGGGCGGTGTGTAC-3′	C2
5′-TGAATG(G/T)GTCCCTGT-3′	MSP-1	Nested PCR (first PCR)
5′-GGAATTCACACCGCCCGTC(A/G)(C/T)TAT-3′	MSP-3
5′-GTTCATTCGCACTACT-3′	MSP-2B	Nested PCR (second PCR)
5′-CCAAGCTTATGCTTAAGTCCAGGGAG-3′	MSP-4B
5′-TGAATG(G/T)GTCCCTGT-3′	MSP-1	Nested PCR (first PCR)
5′-GGAATTCACACCGCCCGTC(A/G)(C/T)TAT-3′	MSP-3
5′-TCACTCGCCGCTACT-3′	MSP-2A	Nested PCR (second PCR)
5′-CCAAGCTTATGCTTAAGT(C/T)(A/C)AA(A/G)GGGT-3′	MSP-4A
5′-CCAGGUTGATUCTGCCUGACG-3′	Mic3U	Nested PCR (first PCR)
5′-TUACCGCGCCUGCUGGCAC-3′	Mic421U
5′-AAGGAGCCTGAGAGATGGCT-3′	Mic266	Nested PCR (second PCR)
5′-CAATTGCTTCACCCTAAGGTC-3′	Eb379
5′-AAGGAGCCTGAGAGATGGCT-3′	Mic266	Nested PCR (second PCR)
5′-CACCCCTTTGCACTCGCACAC-3′	Ec378
5′-AAGGAGCCTGAGAGATGGCT-3′	Mic266	Nested PCR (second PCR)
5′-TGCCCTCCAGTAAATCACAAC-3′	Eh410
5′-AAGGAGCCTGAGAGATGGCT-3′	Mic266	Nested PCR (second PCR)
5′-CCTCCAATCAATCTCGACTC-3′	Ei395
5′-TGCAGTTAAAATGTCCGTAGT-3′	Int530f	5′-TAGCGGCTGACGAAGCTGC-3′
5′-TTTCACTCGCCGCTACTCAG-3′	Int580r	5′-CGGGCAGGAGAACGAGGACGG-3′
		5′-CGGGCAGGAGAACGAGGACGG-3′

Probe designation	Species	Target gene	Amplicon size (bp)	Reference(s)
EB150	E. bieneusi	SSU rRNA	348	145, 149, 260, 262, 381, 395
	E. bieneusi	SSU rRNA	1,265	91, 222, 303
	E. bieneusi	SSU rRNA	607	89
	E. bieneusi	SSU rRNA	443	343
	E. bieneusi	SSU rRNA, ISRa, LSU rRNA	210	357
SI60	E. intestinalis	SSU rRNA	370	81, 148, 149, 260, 262
	E. intestinalis	SSU rRNA	930	91, 222, 303
	E. intestinalis	SSU rRNA	520	90
	E. intestinalis	SSU rRNA	874	343
	E. hellem	SSU rRNA	547	359, 360
	E. cuniculi	SSU rRNA	549	359, 360
	E. bieneusi, E. cuniculi, E. intestinalis, E. hellem	SSU rRNA	250, 268, 270, 279	134
	E. bieneusi, E. cuniculi, E. intestinalis, E. hellem	SSU rRNA	1,200	286
	E. bieneusi	SSU rRNA, ISR, LSU-rRNA	508	190, 191
	E. cuniculi, E. intestinalis, E. hellem	SSU rRNA, ISR, LSU rRNA	289–305	190, 191
	E. bieneusi, E. cuniculi, E. intestinalis, E. hellem	SSU rRNA	410, 419, 421, 433	202
	E. bieneusi	SSU rRNA	132
	E. cuniculi	SSU rRNA	113	202
	E. hellem	SSU rRNA	134	202
	E. intestinalis	SSU rRNA	128	202
	E. cuniculi	SSU rRNA, ISR, LSU rRNA	ca. 1,000	109
	E. hellem
	E. intestinalis

^aISR, intergenic spacer region.