TABLE 3.

VSH and SSCP analysis results for laboratory constructs and three clinical samples previously identified as having mixed HPV-16 variant infections^a

PCR product vol	Sample Id no.	Signal at:				Variants detected by VSH	SSCP analysis resultsb
		Nucleotide position 335		Nucleotide position 350
		C	T	T	G
1×	Group 1	++++	+++	+++++	−	E-350T/Af	s/o multiple
	Group 2	+++	+++	+++	+++++	E-350T/AA	Single
	Group 3	−	++++	+++++	−	E-350T/Af	s/o single
	Group 4	+++++	++	+++++	−	E-350T/Af	Multiple
	OR.1783	++	−	++	+++++	E-350T/AA	Single
	OR.4072	+	++	++	+++++	E-350T/AA	Single
	UK.63550	+++++	−	+++++	+++++	E-G131G/T/AA	Multiple
2×	Group 1	+++++	+++++	+++++	−	E-350T/Af	s/o multiple
	Group 2	++++	+++++	++++	+++++	E-350T/AA	Single
	Group 3	+	+++++	+++++	−	E-350T/Af	s/o single
	Group 4	+++++	+++	+++++	−	E-350T/Af	Multiple
	OR.1783	++++	+++	++++	+++++	E-350T/AA	Single
	OR.4072	++++	++++	++++	+++++	E-350T/AA	Single
	UK.63550	+++++	−	+++++	+++++	E-G131G/T/AA	Multiple
20×	Group 1	+++++	+++++	+++++	−	E-350T/Af	s/o multiple
	Group 2	+++++	+++++	+++++	+++++	E-350T/AA	Single
	Group 3	++	+++++	+++++	−	E-350T/Af	s/o single
	Group 4	+++++	+++++	+++++	−	E-350T/Af	Multiple
	OR.1783	+++++	+++++	+++++	+++++	E-350T/AA	Single
	OR.4072	+++++	+++++	+++++	+++++	E-350T/AA	Single
	UK.63550	+++++	+	+++++	+++++	E-G131G/T/AA	Multiple

^aChemiluminescent signal intensities are as described in footnote b of Table 2. Positive chemiluminescent signals for both possible nucleotides at either one or both nucleotide positions (position 335 or 350) indicated mixed HPV-16 variant infections. 1×, VSH results obtained with a PCR product volume of 4 μl per dot. This is essentially equivalent to the standard volume reported previously (27). 2×, VSH results obtained with a PCR product volume of 8 μl per dot, which is approximately twice the standard volume reported previously (27). 20×, VSH results obtained with a PCR product volume of 80 μl per dot, approximately 20 times the standard volume reported previously (27). Designations of both the major and minor HPV-16 variants present were determined by using probes specific for nucleotide positions 131, 132, 143, 145, 286, and 289 (data not shown), in addition to those listed. 

^bSSCP analyses used a volume of 0.4 μl of PCR product per sample. Results of SSCP analyses are for only a single experiment; however, these results are repeated for each volume to facilitate comparison with VSH results at each volume. SSCP analysis results are defined as follows: Multiple, the presence of multiple HPV-16 variants; s/o multiple, suggestive of the presence of multiple HPV-16 variants; Single, only a single HPV-16 variant was detected; and s/o single, ambiguous results but the presence of only a single HPV-16 variant is suggested.