Tenofovir is a more suitable treatment than entecavir for chronic hepatitis B patients carrying naturally occurring rtM204I mutations

doi:10.3748/wjg.v25.i33.4985

Advanced Search

BPG is committed to discovery and dissemination of knowledge

Home / Archive / Volume 25, Issue 33

This Article

Academic Content and Language Evaluation of This Article

CrossCheck and Google Search of This Article

Academic Rules and Norms of This Article

Citation of this article

Corresponding Author of This Article

Research Domain of This Article

Article-Type of This Article

Open-Access Policy of This Article

Times Cited Counts in Google of This Article

Number of Hits and Downloads for This Article

Total Article Views (6614)

All Articles published online

The chart showing PDF series, WORD series, HTML series, Figures (1-4) series, Tables (1-7) series.

Item

Count

PDF

438

WORD

305

HTML

2875

Figures (1-4)

230

Tables (1-7)

149

Sum=3997

Publishing Process of This Article

The chart showing Browse series, Download series.

Item

Count

Browse

870

Download

1747

Sum=2617

Sep 7, 2019 (publication date) through May 7, 2024

Times Cited of This Article

Times Cited (2)

Journal Information of This Article

Publication Name

World Journal of Gastroenterology

ISSN

1007-9327

Publisher of This Article

Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Observational Study

World J Gastroenterol. Sep 7, 2019; 25(33): 4985-4998
Published online Sep 7, 2019. doi: 10.3748/wjg.v25.i33.4985

Table 1 Primers and LNA probes developed for identification of hepatitis B virus wild type (YMDD) and rtM204I variants (YIDD) by real-time PCR

Primer/probe	Sequence (5'-3') ¹	T_m (°C)²	Target	Channel
Primers (product: 127 bp)
Forward	TGGGCCTCAGTCCGTTTCT	65.4	HBV RT gene
Reverse	TGTACAGACTTGGCCCCCAAWAC	65.2-66.1	HBV RT gene
LNA Probes
YMDD	5' HEX-TAT+A+T+G+G+AT+GAT- 3' IB®FQ	58	YMDD (wild type)	HEX
YIDD	5' 6-FAM-TAT+A+T+T+G+AT+GAT- 3' IB®FQ	53	YIDD	FAM

¹LNA nucleotides are written +A, +C, +T or +G;

²Primer melting temperature was calculated by using LC PDS software V 2.0 and probe melting temperature by https://www.exiqon.com/ls/pages /exiqontmpredictiontool.aspx. T_m: Melting temperature; RT: Reverse transcriptase; HBV: Hepatitis B virus.

Table 2 Measurement of melting temperatures of hepatitis B virus wild type (YMDD) and rtM204I variants (YIDD) by LNA real-time PCR

Sample (copies, number of samples n)	*Measured T_m* (°C) at channel**
	FAM			HEX
	Min	Max	Mean ± SD (number of positives)	Min	Max	Mean ± SD (number of positives)
YIDD positive control DNA (24-2400000, n = 48)	51.3	52.2	51.7 ± 0.2 (48, 100%)	(-)	(-)	(-)
WT positive control DNA (24-2400000, n = 48)	(-)	(-)	(-)	61.9	63.3	62.6 ± 0.4 (46, 95.8%)
YIDD and WT standard mixtures (24-2400000, n = 280)	50.1	52.6	51.5 ± 0.3 (280, 100%)	61.9	64.0	63.0 ± 0.4 (280, 100%)
YIDD and WT standard mixtures (24, n = 56)	49.9	51.9	51.2 ± 0.5 (45, 80.3%)	60.9	63.8	63.0 ± 0.5 (44, 78.6%)
YIDD of clinical samples in duplicate (82-180000, n = 17)	49.6	51.9	51.3 ± 0.5	(-)	(-)	(-)
YMDD of clinical samples in duplicate (65-13000000, n = 397)	(-)	(-)	(-)	61.1	64.0	62.8 ± 0.3

¹The variants were identified by direct sequencing of PCR products. (-) indicates no significant melting temperature; SD: standard deviation; T_m: Melting temperature.

Table 3 Rates of positive detection of hepatitis B virus wild type (YMDD) and rtM204I variant (YIDD) in 410 samples by LNA real-time PCR

Type of detection	No. of samples	Percentage
Clinical samples	410	100%
Identified	403	98.3%
YIDD	17	4.1%
YIDD exclusively	9	2.2%
YIDD + YMDD	3	0.7%
YIDD + YMDD + YIDD with ATA codon¹	1	0.2%
YIDD + YMDD	1	0.5%
YIDD + YMDD	3	0.7%
YMDD	394	96.3%
YMDD exclusively	385	93.9%
YMDD + unknown variant	1	0.2%
Unidentified	7	1.7%
Non-target T_m	2	0.5%
HEX 50.4 °C; YVDD¹	1	0.2%
HEX 57.9 °C; YMDD with Y of TAC codon¹	1	0.2%
No positive signal	5	1.2%

Bold type indicates dominant form.

¹The variants were identified by direct sequencing of PCR products. T_m: Melting temperature.

Table 4 Baseline characteristics of 403 treatment-naïve patients with chronic hepatitis B

Characteristics	Pre-existing rtM204I n = 17	Wild type rtM204 n = 386	P value
Gender (M/F)	14/3	230/156	0.060
Age (yr)	48.8 ± 9.3	43.7 ± 12.6	0.101
HBeAg (positive/negative)	10/7	228/158	0.984
HBV-DNA (log₁₀ IU/mL)	6.33 ± 0.66	6.06 ± 1.77	0.519
qHBsAg (log ₁₀ IU/mL)	3.43 ± 0.30	3.59 ± 0.70	0.334
AST (IU/L)	104.7 ± 40.9	78.7 ± 65.2	0.103
ALT (IU/L)	77.8 ± 29.4	85.4 ± 84.1	0.709
Total bilirubin (mg/dL)	0.93 ± 0.33	0.88 ± 0.60	0.743
Albumin (g/dL)	4.02 ± 0.43	4.14 ± 0.52	0.338
Prothrombin time (INR)	1.07 ± 0.10	1.02 ± 0.16	0.246
Platelet count (× 10³/mm³)	155.7 ± 55.6	181.4 ± 64.4	0.106
FIB-4	4.38 ± 2.33	2.91 ± 2.92	0.041
APRI	1.94 ± 0.98	1.34 ± 1.35	0.068
Significant fibrosis¹ (presence/absence)	12/5	145/241	0.006
Liver cirrhosis (presence/absence)	7/10	80 / 306	0.045
HCC (presence/absence)	5/12	46 / 340	0.034

¹Significant fibrosis was defined as aspartate aminotransferase to platelet ratio index > 1.5 or fibrosis-4 index > 3.25. ALT: Alanine aminotransferase; APRI: Aspartate aminotransferase to platelet ratio index; AST: Aspartate aminotransferase; FIB-4: Fibrosis-4 Index; HCC: Hepatocellular carcinoma; INR: International normalized ratio; qHBsAg: Quantitative hepatitis B surface antigen levels; HBV: Hepatitis B virus.

Table 5 Independent factors for pre-existing rtM204I variants in treatment-naïve chronic hepatitis B patients

	Pre-existing rtM204I (n = 17)	Wild type rtM204 (n = 386)	Univariate			Multivariate
	Pre-existing rtM204I (n = 17)	Wild type rtM204 (n = 386)	OR	95%CI	P value	OR	95%CI	P value
Gender (male)	14 (82.4%)	230 (59.6%)	3.165	(0.895-11.197)	0.074
Age, yr	48.8 ± 9.3	43.7 ± 12.6	1.033	(0.993-1.074)	0.105
HBeAg status (positive)	10 (58.8%)	228 (59.1%)	0.990	(0.369-2.656)	0.984
HBV-DNA, log₁₀ IU/mL	6.33 ± 0.66	6.06 ± 1.77	1.098	(0.827-1.458)	0.519
qHBsAg, log₁₀ IU/mL	3.43 ± 0.30	3.59 ± 0.70	0.555	(0.336-0.916)	0.021
AST, IU/L	104.7 ± 40.9	78.7 ± 65.2	1.005	(0.999-1.011)	0.109
ALT, IU/L	77.8 ± 29.4	85.4 ± 84.1	0.999	(0.992-1.005)	0.709
Total bilirubin, mg/dL	0.93 ± 0.33	0.88 ± 0.60	1.140	(0.522-2.487)	0.742
Albumin, g/dL	4.02 ± 0.43	4.14 ± 0.52	0.655	(0.275-1.559)	0.339
Prothrombin time, INR	1.07 ± 0.10	1.02 ± 0.16	4.625	(0.339-63.020)	0.250
Platelet count, × 10³/mm³	155.7 ± 55.6	181.4 ± 64.4	0.993	(0.985-1.001)	0.108
Significant fibrosis¹	12 (70.6%)	145 (37.6%)	3.989	(1.377-11.553)	0.011	3.397	(1.119-10.319)	0.031
Liver cirrhosis	7 (41.2%)	80 (20.7%)	2.677	(0.988-7.255)	0.053
HCC	5 (29.4%)	46 (11.9%)	3.080	(1.038-9.139)	0.043	1.961	(0.626-6.143)	0.248

Table 6 Independent factors for complete response at 12 mo of antiviral therapy in 232 patients who were treated with nucleos(t)ide analogues

	Complete response (n = 199)	Incomplete response (n = 33)	Univariate			Multivariate
	Complete response (n = 199)	Incomplete response (n = 33)	OR	95%CI	P value	OR	95%CI	P value
Gender (male)	130 (65.3%)	20 (60.6%)	1.225	(0.575-2.610)	0.600
Age, yr	47.3 ± 11.9	43.6 ± 10.9	1.028	(0.996-1.062)	0.087
HBeAg status (positive)	106 (53.3%)	30 (90.9%)	0.114	(0.034-0.386)	< 0.001	0.438	(0.086-2.226)	0.320
HBV-DNA, log₁₀ IU/mL	5.97 ± 1.40	7.69 ± 1.13	0.402	(0.290-0.559)	< 0.001	0.185	(0.083-0.412)	< 0.001
qHBsAg, log₁₀ IU/mL	3.46 ± 0.57	3.89 ± 0.74	0.270	(0.134-0.544)	< 0.001	1.492	(0.501-4.447)	0.473
AST, IU/L	94.6 ± 69.5	83.0 ± 47.2	1.003	(0.997-1.009)	0.361
ALT, IU/L	103.5 ± 92.7	91.6 ± 57.5	1.002	(0.997-1.006)	0.474
Total bilirubin, mg/dL	0.95 ± 0.57	0.76 ± 0.30	2.201	(0.964-5.024)	0.061
Albumin, g/dL	4.06 ± 0.53	4.14 ± 0.44	0.743	(0.351-1.573)	0.438
Prothrombin time, INR	1.05 ± 0.16	1.01 ± 0.10	8.323	(0.444-155.931)	0.156
Platelet count, × 10³/mm³	164.6 ± 63.8	182.6 ± 53.0	0.995	(0.990-1.001)	0.128
Significant fibrosis	105 (52.8%)	13 (39.4%)	1.718	(0.810-3.644)	0.158
Liver cirrhosis	58 (29.1%)	6 (18.2%)	1.851	(0.726-4.720)	0.197
HCC	33 (16.6%)	3 (9.1%)	1.988	(0.573-6.899)	0.279
High genetic barrier	180 (90.5%)	12 (36.4%)	16.579	(7.069-38.882)	< 0.001	82.076	(14.945-450.760)	< 0.001
Pre-existing rtM204I	9 (4.5%)	8 (24.2%)	0.148	(0.052-0.419)	< 0.001	0.014	(0.002-0.096)	< 0.001

ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; INR: International normalized ratio; qHBsAg: Quantitative hepatitis B surface antigen levels; HBV: Hepatitis B virus; HCC: Hepatocellular carcinoma.

Table 7 Treatment responses during nucleos(t)ide analogues in patients with pre-existing rtM204I variants

Outcome	Tenofovir (n = 7)	Entecavir (n = 6)	Low genetic barriers¹ (n = 4)	P value
Reduction of HBV-DNA (log₁₀ IU/mL), mean ± SD
Mo 3	-3.22 ± 0.74	-2.12 ± 0.53²	-2.22 ± 0.40²	0.011
Mo 6	-3.97 ± 0.75	-2.71 ± 0.43²	-2.92 ± 0.51²	0.005
Mo 9	-4.44 ± 0.70	-3.33 ± 0.48²	-3.28 ± 0.35²	0.004
Mo 12	-4.75 ± 0.59	-3.65 ± 0.43²	-3.52 ± 0.60²	0.003
Complete virologic response, cumulative incidence
Mo 12	100%	16.7%²	25%²	0.021

¹The low genetic barriers include lamivudine, adefovir, or combination of lamivudine and adefovir;

²The same number of superscripted indicates non-specific difference between groups. The continuous variables were tested by one-way ANOVA among groups, and categorical variables were tested by Fisher’s exact test. SD: standard deviation; HBV: Hepatitis B virus.

Citation: Choe WH, Kim K, Lee SY, Choi YM, Kwon SY, Kim JH, Kim BJ. Tenofovir is a more suitable treatment than entecavir for chronic hepatitis B patients carrying naturally occurring rtM204I mutations. World J Gastroenterol 2019; 25(33): 4985-4998
URL: https://www.wjgnet.com/1007-9327/full/v25/i33/4985.htm
DOI: https://dx.doi.org/10.3748/wjg.v25.i33.4985