Characteristics of genotypes and resistant mutations within hepatitis B virus (HBV) reverse transcriptase sequences of nucleos(t)ide analogues experienced patients with HBV infection in southern Minnan region

doi:10.3877/cma.j.issn.1674-1358.2019.04.006

Abstract

Abstract:

Objective

To investigate the characteristics of hepatitis B virus (HBV) genotype and reverse transcriptase region (RT) resistance mutation of hepatitis B patients treated with nucleos(t)ide analogues (NAs) in southern Minnan region.

Methods

The K169-M250 nucleotides in HBV RT region of 524 patients with hepatitis B admitted to Liver Diseases Center, the 910th Joint Logistic Support Unit of the People’s Liberation Army of China from January 2012 to December 2017 were analyzed by direct sequencing of PCR products. The patients were followed up after antiviral therapy, and the levels of HBV e antigen (HBeAg), HBV DNA, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected, respectively.

Results

Among the enrolled 524 patients, 293 cases (55.92%) were HBV genotype B and 231 cases (44.08%) were HBV genotype C. The sex, HBeAg status, course of disease, selection of NAs drug, mutation rate of drug resistance after NAs application, period of NAs application and serum HBV DNA level, levels of ALT and AST were not significantly different (all P > 0.05). However, the age of patients with HBV genotype C (t = 6.486, P < 0.001) and mutation rates of NAs related drug resistance (64.94% vs. 51.88%; χ² = 8.493, P = 0.003) were significantly higher than those of patients with HBV genotype B, with significant differences. Among the 524 patients, the detection rate of NAs related drug resistance mutation was 57.63% (302/524), and the frequency of mutation at site 204 was the highest, followed by 180, 181 and 229 sites. Ten mutations with M204 mutation were found, including 169, 173, 180, 200, 202, 207, 214, 237, 245 and 250 sites. The mutation rates of patients with HBV genotype C at 9 sites (180, 181, 184, 191, 200, 221, 224, 229 and 238 sites) were significantly higher than those of patients with HBV genotype B, with significant differences (all P < 0.05). The mutation rates of HBV B-type patients at 236 and 250 sites were significantly higher than those of C-type patients (χ² = 5.867, P = 0.015; χ² = 4.226, P = 0.040). The mutation associated with M204 also showed different tendency in HBV genotype B and C: M204I mutation was more likely to occur in patients with genotype B, and L180 + M204I, T184 + L180 + M204V were more likely to be present in patients with genotype C.

Conclusion

Under the pressure of NAs drug selection, HBV genotype B and C infection showed different characteristics of drug resistances mutation.

Key words: Hepatitis B virus, Genotype, Resistance mutation, Nucleoside/nucleotide analogues

Xiaoman Zhang, Tao Xu, Zhengju Xu. Characteristics of genotypes and resistant mutations within hepatitis B virus (HBV) reverse transcriptase sequences of nucleos(t)ide analogues experienced patients with HBV infection in southern Minnan region[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2019, 13(04): 293-299.

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References 27

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位点类型	个数	位点名称
主要耐药变异位点	8	I169T、A181T/V、T184A/C/F/G/I/L/M/S、A194T、S202C/G/I、M204I/V、N236T、M250I/L/V
次要/代偿性耐药变异位点	3	L80I/V、V173L、L180M
假定耐药变异位点	26	S53N、T54N、L82M、V84M、S85A、I91L、Y126C、T128N、N139D^a、W153Q、F166L、V191I、A200V、V207I、S213T、V214A、Q215P/S、L217R、E218D、F221Y、L229G/V/W、I233V、P237H、N238D/S/T、Y245H、S/C256G
治疗前耐药变异位点	6	T38A、Y124H、D134E、N139K/H^a、I224V、R242A

位点类型	个数	位点名称
主要耐药变异位点	8	I169T、A181T/V、T184A/C/F/G/I/L/M/S、A194T、S202C/G/I、M204I/V、N236T、M250I/L/V
次要/代偿性耐药变异位点	3	L80I/V、V173L、L180M
假定耐药变异位点	26	S53N、T54N、L82M、V84M、S85A、I91L、Y126C、T128N、N139D^a、W153Q、F166L、V191I、A200V、V207I、S213T、V214A、Q215P/S、L217R、E218D、F221Y、L229G/V/W、I233V、P237H、N238D/S/T、Y245H、S/C256G
治疗前耐药变异位点	6	T38A、Y124H、D134E、N139K/H^a、I224V、R242A

临床特征		例数	HBV B型（293例）	HBV C型（231例）	统计量	P值
性别[例（%）]		?	?	?	χ² = 0.493	0.483
?	男	430	244（56.74）	186（43.26）	?	?
?	女	94	49（52.13）	45（47.87）	?	?
HBeAg [例（%）]		?	?	?	χ² = 0.010	0.919
?	（+）	406	227（55.96）	179（44.04）	?	?
?	（－）	118	66（53.85）	52（46.15）	?	?
病程[例（%）]		?	?	?	χ² = 4.821	0.090
?	CHB	428	250（57.94）	178（42.06）	?	?
?	LC	68	27（39.71）	41（60.29）	?	?
?	PHC	28	11（39.29）	17（60.71）	?	?
NAs药物选择[例（%）]		?	?	?	χ² = 6.030	0.197
?	LAM	79	46（57.50）	33（42.50）	?	?
?	LDT	33	21（64.71）	12（35.29）	?	?
?	ADV	65	28（42.42）	37（57.58）	?	?
?	ETV	114	77（67.54）	37（32.46）	?	?
?	联合用药	233	127（54.62）	106（45.38）	?	?
NAs相关耐药突变率[例（%）]		302	152（50.33）	150（49.67）	χ² = 0.001	0.981
不同NAs治疗耐药突变率[例（%）]		?	?	?	χ² = 7.383	0.117
?	LAM	55	28（50.91）	27（49.09）	?	?
?	LDT	24	16（66.67）	8（33.33）	?	?
?	ADV	32	13（40.62）	19（59.38）	?	?
?	ETV	43	28（65.12）	15（34.88）	?	?
?	联合用药	148	72（48.65）	76（51.35）	?	?
用药时间（ ± s，月）		524	42.00 ± 31.05	41.00 ± 31.83	t = 0.207	0.835
年龄（ ± s，岁）		524	36.94 ± 11.47	43.91 ± 13.10	t = 6.486	< 0.001
HBV DNA （ ± s，log₁₀ IU/ml）		524	5.35 ± 1.49	5.39 ± 1.56	t = 0.293	0.770
ALT（ ± s，U/L）		524	99.30 ± 255.48	163.13 ± 269.27	t = 0.346	0.730
AST（ ± s，U/L）		524	128.40 ± 193.46	116.17 ± 271.46	t = 0.646	0.519

临床特征		例数	HBV B型（293例）	HBV C型（231例）	统计量	P值
性别[例（%）]		?	?	?	χ² = 0.493	0.483
?	男	430	244（56.74）	186（43.26）	?	?
?	女	94	49（52.13）	45（47.87）	?	?
HBeAg [例（%）]		?	?	?	χ² = 0.010	0.919
?	（+）	406	227（55.96）	179（44.04）	?	?
?	（－）	118	66（53.85）	52（46.15）	?	?
病程[例（%）]		?	?	?	χ² = 4.821	0.090
?	CHB	428	250（57.94）	178（42.06）	?	?
?	LC	68	27（39.71）	41（60.29）	?	?
?	PHC	28	11（39.29）	17（60.71）	?	?
NAs药物选择[例（%）]		?	?	?	χ² = 6.030	0.197
?	LAM	79	46（57.50）	33（42.50）	?	?
?	LDT	33	21（64.71）	12（35.29）	?	?
?	ADV	65	28（42.42）	37（57.58）	?	?
?	ETV	114	77（67.54）	37（32.46）	?	?
?	联合用药	233	127（54.62）	106（45.38）	?	?
NAs相关耐药突变率[例（%）]		302	152（50.33）	150（49.67）	χ² = 0.001	0.981
不同NAs治疗耐药突变率[例（%）]		?	?	?	χ² = 7.383	0.117
?	LAM	55	28（50.91）	27（49.09）	?	?
?	LDT	24	16（66.67）	8（33.33）	?	?
?	ADV	32	13（40.62）	19（59.38）	?	?
?	ETV	43	28（65.12）	15（34.88）	?	?
?	联合用药	148	72（48.65）	76（51.35）	?	?
用药时间（ ± s，月）		524	42.00 ± 31.05	41.00 ± 31.83	t = 0.207	0.835
年龄（ ± s，岁）		524	36.94 ± 11.47	43.91 ± 13.10	t = 6.486	< 0.001
HBV DNA （ ± s，log₁₀ IU/ml）		524	5.35 ± 1.49	5.39 ± 1.56	t = 0.293	0.770
ALT（ ± s，U/L）		524	99.30 ± 255.48	163.13 ± 269.27	t = 0.346	0.730
AST（ ± s，U/L）		524	128.40 ± 193.46	116.17 ± 271.46	t = 0.646	0.519

突变位点	突变例数	伴随M204I	伴随M204V	伴随M204I/V
I169	2	1（50.00）	1（50.00）	0（0.00）
V173	6	3（50.00）	2（33.33）	1（16.67）
L180	109	20（18.35）	83（76.15）	6（5.50）
A200	12	4（33.33）	8（66.67）	0（0.00）
S202	24	0（0.00）	24（100.00）	0（0.00）
V207	9	4（44.44）	4（44.44）	1（11.12）
V214	2	2（100.00）	0（0.00）	0（0.00）
P237	3	1（33.33）	2（66.67）	0（0.00）
Y245	1	1（100.00）	0（0.00）	0（0.00）
M250	11	7（63.64）	4（36.36）	0（0.00）

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