TABLE 2.

Semiquantitative dot blot results for 14 clinical samples containing multiple HPV-16 variants^a

Sample no.	Signal atb:												Variants present by VSHc
	Nucleotide position 335						Nucleotide position 350
	1×		2×		20×		1×		2×		20×
	C	T	C	T	C	T	T	G	T	G	T	G
OR.5392	+++	−	+++++	−	+++++	−	−	++++	+	+++++	+++	+++++	E-G/T
OR.6428	++++	−	+++++	−	+++++	−	−	++++	+++	++++++	+++++	+++++	E-G/T
OR.9887	++++	−	+++++	−	+++++	−	−	++++	+	+++++	++	+++++	E-G/T
UK.41484	++++	−	+++++	−	+++++	−	−	++++	−	+++++	++	+++++	E-G/T
UK.11902	++++	−	+++++	−	+++++	−	++++	−	+++++	+	+++++	+++	E-G/T
OR.3127	++++	−	+++++	−	+++++	−	++++	−	+++++	++	+++++	+++	E-G/T
OR.1783d	++	−	++++	+++++	+++++	+++++	+++	++++	+++++	+++++	+++++	+++++	E/NE
OR.4072d	+	++	++++	+++++	+++++	+++++	++	++++	+++++	+++++	+++++	+++++	E/NE
WA.166	−	++++	+	+++++	+++	+++++	−	++++	++	+++++	+++	+++++	E/NE
WA.74	−	++++	++	+++++	++++	+++++	−	++++	++++	+++++	+++++	+++++	E/NE
WA.101	−	++++	+	+++++	++	+++++	−	++++	+	+++++	++	+++++	E/NE
WA.201	+	++++	++	+++++	++++	+++++	−	++++	++++	+++++	++++	+++++	E/NE
WA.153	−	+++	−	+++++	++	+++++	−	++++	−	+++++	++	+++++	E/NE
WA.188	−	+++	+	+++++	++	+++++	−	++++	+	+++++	+	+++++	E/NE
UK.40090	++++	−	+++++	−	+++++	−	−	++++	++	+++++	++++	+++++	E-G/T
WA.40084A	++++	−	+++++	−	+++++	−	−	++++	−	+++++	++	+++++	E-G/T
WA.272d	++++	++	+++++	++++	+++++	+++++	++++	+++	+++++	+++++	+++++	+++++	E/NE

^aAs described in Materials and Methods, only probe pairs for HPV-16 nucleotide positions 335 (C versus T) and 350 (T versus G) were used in these particular studies. Thus, HPV-16 variant class designations could be defined only as either European (E) or non-European (NE). 

^bThe relative volumes of resultant amplification reaction mixtures are designated 1×, 2×, and 20×. Chemiluminescent signal intensities recorded on X-ray film are as follows: +++++, very strong hybridization signal; ++++, a strong signal; +++, a moderate signal; ++, a weak signal; +, a very weak signal; and −, no detectable signal. Positive chemiluminescent signals for both possible nucleotides at either one or both nucleotide positions (position 335 or 350) indicated mixed HPV-16 variant infections (i.e., sample WA.201 gave signal intensities of + with the 335C probe and ++++ with the 335T probe with a PCR product volume of 1×, indicating the potential presence of both European and non-European variants). These results were confirmed with the 2× and 20× PCR product volumes. 

^cThe variant designations are E-G/T (European 350G mixed with European 350T) and E/NE (European variant mixed with a non-European variant, i.e., Af-1, Af-2, AA, or NA-1). 

^dSamples previously identified as having mixed HPV-16 variant infections by VSH and included in this study as controls.