Identification of reliable biomarkers of human papillomavirus 16 methylation in cervical lesions based on integration status using high-resolution melting analysis

Liu, Lu; Ying, Chunmei; Zhao, Zhen; Sui, Long; Zhang, Xinyan; Qian, Chunyan; Wang, Qing; Chen, Limei; Guo, Qisang; Wu, Jiangnan

doi:10.1186/s13148-018-0445-8

Table 2 The HRM primers were designed with Methyl Primer software to screen for methylation of a total of 61 CpG sites in the HPV genome, based on the results of the integration experiments, in order to avoid breakpoints in each sample

From: Identification of reliable biomarkers of human papillomavirus 16 methylation in cervical lesions based on integration status using high-resolution melting analysis

Primer	Sequence (5′–3′)	CpG sites	Size (bp)
E2BSs	F-AATTTATGTATAAAATTAAG	nt31, nt37, nt43,	106
E2BSs	R-CCTAAAACATTACAATTCTCTT	nt52, nt58	106
E2	F-TTGAAATTATTAGGTAGTATTTGGTT	nt3412, nt3415, nt3417, nt3433, nt3461, nt3462
E2	R-TAATYGTCTATATTTCTTYGATG	nt3412, nt3415, nt3417, nt3433, nt3461, nt3462	90
L1-1	F-GTTTTTTGTAGATTTAGATTAGTTTTTTT	nt7034, nt7091, nt7136, nt7145	187
L1-1	R-AAACAACACTAATTCAACATACATACAA	nt7034, nt7091, nt7136, nt7145	187
L1-2	F-TGTTAGAATTATATGGCGATAGTT	nt6367, nt6389, nt6457	124
L1-2	R-CCTTTAATATATAAATCGTCTAATACATT	nt6367, nt6389, nt6457	124
L1-3	F-TAATTTAGATATATAGCGGTTGGT	nt5927, nt5963	81
L1-3	R-CTAATACCCACACCTAATAACTAA	nt5927, nt5963	81
L1-4	F-GAGTATAGGGTTATAATAATGGT	nt6581, nt6650	109
L1-4	R-AAATAAATATAACAACACATAATAACAT	nt6581, nt6650	109
L1-5	F-TTAAGGAGTATTTACGATATGGG	nt6731, nt6796	199
L1-5	R-ACAATTACCTAAATTACAAACCTATA	nt6731, nt6796	199
L1-6	F-TTTGATATATTTATTAATATAATTGATTA	nt5600, nt5606, nt5609, nt5615	142
L1-6	R-CYTTTACYTCYTTTTCYTAACATATA	nt5600, nt5606, nt5609, nt5615	142
L1-7	F-TGTAAGTACGGATGAATATGTT	nt5709, nt5726	81
L1-7	R-AAATATCCAACTACAAATAATCTAAA	nt5709, nt5726	81
L2-1	F-AATAAATTGTTATTATTTAATAATGCGAT	nt4240, nt4249, nt4261, nt4270, nt4277	120
L2-1	R-ATAATATCAAATAAACATATACCTACCTA	nt4240, nt4249, nt4261, nt4270, nt4277	120
L2-2	F-TTTGATATTATATTTAAGGTTGAAGGT	nt4427, nt4437, nt4441	125
L2-2	R-CCAATACGTCCGCCTATAC	nt4427, nt4437, nt4441	125
L2-3	F-ATAGTAATTAGTAGTATATTTATATTAGG	nt4894, nt4906, nt4924	90
L2-3	R-TACAACTTTAACCTATTATATTATACG	nt4894, nt4906, nt4924	90
L2-4	F-TTAGGCGTATTGGTATTAGGT	nt5128, nt5173, nt5179	101
L2-4	R-ATCATAATAATAATATACCTTAACACCT	nt5128, nt5173, nt5179	101
L2-5	F-TTTTATAATTTYGGTAGGATTTGTA	nt5378	150
L2-5	R-CTTAATCAATTATATTAATAAATATATCAA	nt5378	150
E6	F-TTGTTAATTAGGTGTATTAATTGTTAA	nt494, nt502, nt506, nt539	179
E6	R-TATATCTCCATACATAATTACAACTAA	nt494, nt502, nt506, nt539	179
E7	F-GATAAGTAGAATCGGATAGAGTT	nt701, nt752, nt757, nt765, nt780, nt789	137
E7	R-CCTAATATACCCATTAACAAATCTT	nt701, nt752, nt757, nt765, nt780, nt789	137
Clone-LCR	F-CCTACTAATTGTGTTGTGGT		689
Clone-LCR	R-CTCCTGTGGGTCCTGAA		689
Clone-E6	F-GGAACAACATTAGAACAGCA		231
Clone-E6	R-GCAATGTAGGTGTATCTCCA		231
Clone-E7	F-GCAACCAGAGACAACTGA		231
Clone-E7	R-GGGCACACAATTCCTAGT		231
Clone-E2	F-AGCAACGAAGTATCCTCTC		187
Clone-E2	R-TGTCCACTGAGTCTCTGT		187
Clone-L2	F-CCTCTGCGTTTAGGTGTT		1504
Clone-L2	R-GAAGGAGCTTGGTCAGTTAT		1504
Clone-L1	F-TTCCAGGGTCTCCACAATA		1693
Clone-L1	R-GCACATACAAGCACATACAA		1693

Primers for high-resolution melting PCR for determination of human papillomavirus 16 methylation and construction of standard curves

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ISSN: 1868-7083

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