Table 1.

Novel quantitative PCR assays for non-vaccine serotypes.

Serotype(s)		Sequencea	Target region	Accession No.	Position	Size (bp)	Limit of detection genome equivalents (fg)	Concentration (nM)	Reaction efficiency (avg)
10CF	Forward	CGAGTTATGGATGTTCTTATTGGC	wcjG	CR931651	4808–4831	137	21.4 (50)	400	103.4
	Reverse	CCCAACCCCACTCTGTATTG			4925–4944			400
	Probe	ACAGGGCAAGACTGTGAATATTGTTCCA			4837–4864			300
11BC	Forward	TCAAATTTGGCGTATTGCTTATCA	wzy	CR931654	10 998–11 021	106	2.1 (5)	400	102.8
	Reverse	TGATTATGAGCATAGTTGATCCCC			11 103–11 126			400
	Probe	TCCGTGGCAAGATTCTGGTGCTAAG			11 079–11 103			200
16A	Forward	GCTAGCAGGAACTTTTCTAGGG	wcxR	CR931667	6675–6696	132	21.4 (50)	200	92.9
	Reverse	TCCCTGTCCAAATCCGAAAC			6787–6806			200
	Probe	CCCACGGGATGAATCCATTATGGCG			6703–6727			200
17A	Forward	TGATTATGTCATTCGATTGCTTGG	wzy	CR931669	13 895–13 918	112	2.1 (5)	400	93.5
	Reverse	AAATCCTAAAATTCCTGTTTGAAAAGC			13 980–14 006			400
	Probe	ATTATGGGCGTGGGTTACCGTAGG			13 941–13 964			200
17F	Forward	TGCTTTTGTGGGTAGGACAAG	wzx	CR931670	17 361–17 381	130	21.4 (50)	400	97.8
	Reverse	TTATCCCATAAACCTGAGGCG			17 470–17 490			400
	Probe	TGCAGGTGATATGCGGAGCCAAT			17 440–17 462			200
19C	Forward	AATGGTTTTCAGATTACTTGATAGCTC	wchU	CR931677	19 294–19 320	115	2.1 (5)	400	95.7
	Reverse	CGTTCCTTATGAGAGTGGTCAAG			19 386–19 408			400
	Probe	TGTTCCTGCCCCCACATAATGAACT			19 343–19 367			200
24BF	Forward	TCTGAAAGTAATTAGTAAGATTAACGGAAG	wzy	CR931688	15 038–15 067	133	2.1 (5)	400	101.5
	Reverse	TCCATCTACTTTTAAAATAGCTCCAAC			15 144–15 170			400
	Probe	CCACAGTCCCAAAATTGTCAGCAACC			15 085–15 110			200
28AF	Forward	CAACTACAGGTATTTTTGATATCGGAG	wcxP	CR931692	12 047–12 073	141	2.1 (5)	400	96.7
	Reverse	GTTTACTACGTTTGTGAAGCGC			12 166–12 187			400
	Probe	AGAAAATAGTAGGTTGATTGGCGGTGCT			12 075–12 102			200
32AF	Forward	GTACTTCCTGTTCTAGGCTTGG	wzy	CR931696	12 471–12 492	134	2.1 (5)	400	101.7
	Reverse	CCCAGAGGAAAATAGCGTCTC			12 584–12 604			400
	Probe	TTGTTCAAACCCAACCACTGCTCC			12 492–12 515			200
33C	Forward	CAGAGACAGTTTCAGCAAATCTTAG	wzy	CR931700	11 270–11 294	147	2.1 (5)	400	103
	Reverse	AGCCTACACCTCTTATAAACGTTG			11 393–11 416			400
	Probe	CCGTGTCCTATCCACAAACTTGTCTTCC			11 327–11 354			200
48	Forward	CAGGTTTTGCTTCATATGGGAG	wzy	CR931722	13 043–13 064	133	2.1 (5)	400	94.9
	Reverse	ATCGGCCAAAAGTTATCATTAGC			13 153–13 175			400
	Probe	CGCTGCTTATGTGTATTACTCTCCCCTG			13 068–13 095			200

^aProbes were labeled at 5΄ with FAM (6-carboxyfluorescein) and at 3΄ with BHQ1 (Black Hole Quencher-1).