De Weggheleire Anja, De Baetselier Irith, An Sokkab, Goletti Sylvie, Suin Vanessa, Thai Sopheak, Francque Sven, Crucitti Tania, Lynen Lutgarde, Van Gucht Steven, Kabamba Benoît Mukadi
Clinical Sciences Department, Institute of Tropical Medicine Antwerp, Antwerp, Belgium.
Infectious Diseases Department, Sihanouk Hospital Center of Hope, Phnom Penh, Cambodia.
Infect Dis Ther. 2020 Sep;9(3):657-667. doi: 10.1007/s40121-020-00304-7. Epub 2020 May 30.
We aim to report on results and challenges of different methods used for hepatitis C (HCV) genotyping in a Cambodian HCV/HIV coinfection project.
Samples of 106 patients were available. HCV genotyping was initially (63 samples) done by the LightPower Taqman real-time PCR method (Viet A Corp.) and quality controlled using the Versant 2.0 line probe assay (Siemens Healthcare). Next, following interim quality control results, all 106 samples were (re)genotyped with Versant 2.0, complemented with 5'UTR/core sequencing for uninterpretable/incomplete Versant results.
Using Versant, 103 (97.2%) of the 106 HCV-coinfected patients had an interpretable genotype result: 1b (50.5%), 6 non-a/non-b (30.1%), 1a (6.8%), 6a or b (4.9%), 2 (3.9%), 1 (2.9%) and 3 (1.0%). For 16 samples that were interpreted as genotype 1 or 1b per Versant's current instructions, it could not be excluded that it concerned a genotype 6 infection as the core region line patterns on the Versant test strip were unavailable, inconclusive or atypical. Upon sequencing, seven of these were genotyped as 1b and nine as genotype 6. Combining Versant and sequencing results, a definitive genotype was assigned in 104 patients: 1b (44.2%), 6 non-a/non-b (39.4%), 1a (6.7%), 6a or b (4.8%), 2 (3.8%) and 3 (1.0%). Genotyping by LightPower and Versant was discordant for 23 (of 63) samples. The LightPower assay misclassified all genotype 6 non-a/non-b samples as genotype 1, which indicates that this assay is only using 5'UTR information.
HCV genotype 1b and genotype 6 non-a/non-b were most common. With Versant 2.0 (using 5'UTR and core information), genotype classification (1 or 6) remained inconclusive in 15% of samples. The locally available method (LightPower assay) failed to identify genotype 6 non-a/non-b, which highlights that methods using 5'UTR information only should not be used in Cambodia. Regional/national guidelines should be explicit about this.
This study was performed as part of a larger cross-sectional study on the burden of hepatitis C coinfection in HIV patients in Cambodia (Clinical.trials.gov: HCV-Epi NCT02361541).
我们旨在报告柬埔寨丙型肝炎病毒（HCV）/人类免疫缺陷病毒（HIV）合并感染项目中用于HCV基因分型的不同方法的结果和挑战。
有106例患者的样本可供使用。HCV基因分型最初（63个样本）采用LightPower Taqman实时荧光定量PCR方法（Viet A公司）进行，并使用Versant 2.0线性探针检测法（西门子医疗）进行质量控制。接下来，根据中期质量控制结果，所有106个样本均采用Versant 2.0重新进行基因分型，并对无法解释/不完整的Versant结果补充5'非翻译区/核心区测序。
使用Versant检测，106例HCV合并感染患者中有103例（97.2%）获得了可解释的基因型结果：1b型（50.5%）、6型非a/b型（30.1%）、1a型（6.8%）、6a或b型（4.9%）、2型（3.9%）、1型（2.9%）和3型（1.0%）。对于按照Versant当前说明被解释为1型或1b型的16个样本，由于Versant测试条上的核心区条带模式不可用、不确定或不典型，不能排除其为6型感染。测序后，其中7个样本被基因分型为1b型，9个样本为6型。结合Versant和测序结果，104例患者被确定了明确的基因型：1b型（44.2%）、6型非a/b型（39.4%）、1a型（6.7%）、6a或b型（4.8%）、2型（3.8%）和3型（1.0%）。LightPower和Versant基因分型对63个样本中的23个结果不一致。LightPower检测将所有6型非a/b型样本错误分类为1型，这表明该检测仅使用了5'非翻译区信息。
HCV 1b型和6型非a/b型最为常见。使用Versant 2.0（使用5'非翻译区和核心区信息）时，15%的样本基因型分类（1型或6型）仍不明确。当地可用的方法（LightPower检测）未能识别6型非a/b型，这突出表明仅使用5'非翻译区信息的方法不应在柬埔寨使用。地区/国家指南应对此予以明确说明。
本研究是柬埔寨关于HIV患者丙型肝炎合并感染负担的一项更大规模横断面研究的一部分（Clinical.trials.gov：HCV-Epi NCT02361541）。
