Real-Life Diagnostic Accuracy and Clinical Utility of Hepatitis B Virus (HBV) Nucleic Acid Testing Using the GeneXpert Point-of-Care Test System from Fresh Plasma and Dry Blood Spot Samples in The Gambia
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Participants
2.1.1. Sample Collection
2.1.2. DBS Optimization: Limit of Detection
2.2. Plasma
2.3. TAT and GeneXpert Mapping
2.4. Data Analysis
3. Results
3.1. DBS Optimization: Limit of Detection (LOD)
3.2. Plasma and DBS HBV Viral Load—Group 1
3.3. Plasma HBV Viral Load—Group 2
3.4. GeneXpert Against in-House qPCR
3.5. GeneXpert Against COBAS TaqMan Assays
3.6. In-House qPCR Versus COBAS TaqMan for Plasma Viral Load
3.7. Analysis Using Plasma GeneXpert Against DBS GeneXpert
3.8. Turn-Around Time, GeneXpert Mapping and Real-World Challenges
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Batch 5 | ||
---|---|---|
Sample 1 = 50 μL—3 Punches (650 μL Elution) | DBS Viral Load | Plasma Viral Load |
EG5894 | 1.73 × 103 IU/mL log 3.24 | 5.47 × 104 IU/mL log 4.74 |
EG5892 | <10 IU/mL log 1.00 | 276 IU/mL log 2.44 |
EG5102 V2 | HBV NOT DETECTED | <10 IU/mL log 1.00 |
EG5095 V2 | HBV NOT DETECTED | HBV NOT DETECTED |
Sample 2 = 75 μL—4 Punches (750 μL Elution) | ||
EG5894 | 1.75 × 103 IU/mL log 3.24 | 5.47 × 104 IU/mL log 4.74 |
EG5892 | <10 IU/mL log 1.00 | 276 IU/mL log 2.44 |
EG5102 | <10 IU/mL log 1.00 | <10 IU/mL log 1.00 |
EG5095 | HBV NOT DETECTED | HBV NOT DETECTED |
Sample 3 = 100 μL—6 Punches (1000 μL Elution) | ||
EG5894 | 2.07 × 103 IU/mL log 3.32 | 5.47 × 104 IU/mL log 4.74 |
EG5892 | 12 IU/mL log 1.08 | 276 IU/mL log 2.44 |
EG5102 | HBV NOT DETECTED | <10 IU/mL log 1.00 |
EG5095 | HBV NOT DETECTED | HBV NOT DETECTED |
GeneXpert HBV Viral Load Plasma (No. of Plasma) | GeneXpert HBV DBS Viral Load (No. of DBS) | |
---|---|---|
Not detected | 3 | 8 |
<10 IU/mL | 4 | 15 |
≥20 IU/mL | 51 | 33 |
Total | 58 | 56 |
Variables | Number (%) | |
---|---|---|
Sex | Men | 202 (71%) |
Women | 81 (29%) | |
Age (years) | 18–29 | 23 (8%) |
30–39 | 120 (43%) | |
40–49 | 83 (29%) | |
≥50 | 57 (20%) | |
Disease status (by USA) | No apparent liver disease | 261 (92%) |
Cirrhosis | 14 (5%) | |
HCC | 1 (1%) | |
Fatty liver disease | 7 (2%) |
GeneXpert | In-House PCR | COBAS TaqMan | ||
---|---|---|---|---|
Undetectable HBV DNA (n = 167) | Detectable HBV DNA (n = 116) | Undetectable HBV DNA n = 137 | Detectable HBV DNA n = 117 | |
Undetectable | 39 (23%) | 0 (0%) | 0 (0%) | 0 (0%) |
Detectable | 120 (72%) | 109 (94%) | 129 (85.3%) | 110 |
Error | 4 (2%) | 3 (3%) | 4 (2%) | 3 (3%) |
Invalid | 4 (2%) | 4 (3%) | 4 (2%) | 4 (3%) |
Plasma Samples | |||||
---|---|---|---|---|---|
Undetectable (n = 14) | Detectable (n = 73) | Error (n = 2) | Invalid (n = 2) | ||
DBS samples | Undetectable (n = 18) | 5 (36%) | 11 (15%) | 1 (50%) | 1 (50%) |
Detectable (n = 37) | 1 (7%) | 34 (47%) | 1 (50%) | 1 (50%) | |
Error (n = 3) | 0 (0%) | 3 (4%) | 0 (0%) | 0 (0%) | |
Invalid (n = 33) | 8 (57%) | 25 (34%) | 0 (0%) | 0 (0%) |
Median TAT | GeneXpert | qPCR |
---|---|---|
From HBsAg testing to enrollment to the study | 8 days (IQR (8–12.5)) | N/A |
Sample collection to HBV DNA testing completion | 138 min (IQR 113–179) | 83 days (28–122) |
HBV DNA results to treatment initiation | 6 days (IQR 5.5–6.5) | 18 days (16–37) |
Regions | No. Health Facilities | GeneXpert Module (s) and Cartridge No | Purpose (Assays) | Funders | Cartridge Loss (Error or Invalid)/Average Per Week |
---|---|---|---|---|---|
West Coast | 3 | 4 (1), 16 (2) | HIV, TB, and early infant diagnosis | Global Fund, USA Army | 6 |
Greater Banjul | 8 | 4 (8), 16 (2) and infinity (1) | COVID-19, HIV, TB, HBV, early infant diagnosis, and host response 3-gene research | Global Fund | 6 |
Central River | 2 | 4 (2) | COVID-19, HIV, TB, and early infant diagnosis | Global Fund | 10 |
Upper River | 1 | 4 (1) | COVID-19, HIV, TB | Global Fund | 5 |
North Bank | 2 | 4 (2) | COVID-19, HIV, TB and early infant diagnosis | Global Fund, UNICEF | 5 |
Lower River | 1 | 4 (1) | HIV, TB and early infant diagnosis | Global Fund | 5 |
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Ceesay, A.; Drammeh, S.; Ndow, G.; Jallow, A.O.A.; Nyang, H.; Bittaye, B.; Mendy, F.S.; Secka, O.; D’Alessandro, U.; Shimakawa, Y.; et al. Real-Life Diagnostic Accuracy and Clinical Utility of Hepatitis B Virus (HBV) Nucleic Acid Testing Using the GeneXpert Point-of-Care Test System from Fresh Plasma and Dry Blood Spot Samples in The Gambia. Microorganisms 2024, 12, 2273. https://doi.org/10.3390/microorganisms12112273
Ceesay A, Drammeh S, Ndow G, Jallow AOA, Nyang H, Bittaye B, Mendy FS, Secka O, D’Alessandro U, Shimakawa Y, et al. Real-Life Diagnostic Accuracy and Clinical Utility of Hepatitis B Virus (HBV) Nucleic Acid Testing Using the GeneXpert Point-of-Care Test System from Fresh Plasma and Dry Blood Spot Samples in The Gambia. Microorganisms. 2024; 12(11):2273. https://doi.org/10.3390/microorganisms12112273
Chicago/Turabian StyleCeesay, Amie, Sainabou Drammeh, Gibril Ndow, Alpha Omar A. Jallow, Haddy Nyang, Baboucarr Bittaye, Francis S. Mendy, Ousman Secka, Umberto D’Alessandro, Yusuke Shimakawa, and et al. 2024. "Real-Life Diagnostic Accuracy and Clinical Utility of Hepatitis B Virus (HBV) Nucleic Acid Testing Using the GeneXpert Point-of-Care Test System from Fresh Plasma and Dry Blood Spot Samples in The Gambia" Microorganisms 12, no. 11: 2273. https://doi.org/10.3390/microorganisms12112273