Inconsistent virus DNA quantification results in clinical hepatitis B and the verification by genotyping and sequencing: a case report

Hepatitis B virus (HBV) infection is the leading cause of hepatocellular carcinoma (HCC). Approximately 700,000 people die of HCC each year worldwide and HCC is thus the third leading cause of cancer related deaths [1]. Despite the HBV vaccine program established in the early 1990s, an epidemic of HBV infection in China still causes a rigorous public health problem [2]. Given the high risk of developing liver cirrhosis andHCCafter chronicHBV infections, there is a critical need for screening, diagnosis and monitoring of HBV [3]. In addition to tests for HBV serological markers, real-time PCR based HBV DNA quantification has been widely used in clinical practice to predict disease activity and monitor virus treatment response [4]. We herein present a casewith opposite results of HBVDNAquantification between the Sansure method and the Roche method using the same patient serum. A 52-year old male with a history of HBV infection and cirrhosis for seven years was admitted to Qianfoshan hospital on February 9th, 2019. The patient was given a diagnosis of HCC. His serum HBV DNA quantification results (Sansure HBVDNAquantification kit, China; ABI 7300 Plus instrument, USA) revealed that the virus concentrationwas 1.28E + 05 IU/mL. Three days later, a second serum of this patient was drawn and examined by Roche (COBAS AmpliPrep/COBAS TaqMan48, HBV Test v2.0, USA) HBV DNA quantification test due to a temporary shortage of Sansure kits and the result was <2.00E + 01 IU/mL. The inter-laboratory comparison between Sansure method and Roche method was performed every three months, and there were no inconsistent results observed ever. On the other hand, it was uncommon to see a dramatic decline of HBV copies in clinic. The sample was further confirmed to have neither hemolytic nor lipid interference, and the patient was overnight fasting before blood collection. The instruments used for quantification were under quality inspection and maintained daily. The internal controls in experiments and the amplification curves were normal (Figure 1). To eliminate the possibility of operating error, each determination method was repeated twice more. Resultswith the Sansuremethodwere consistentwith the first test (1.08E + 05 IU/mL, 1.13E + 05 IU/mL), and results with the Roche method were consistent with the prior Roche results (undetermined, undetermined). To help analyze the results, the patient’s related clinical and laboratory parameters were reviewed. The patient has a seven year history of HBV infection and has been taking anti-HBV drugs. The HBsAg level regularly monitored two months ago was 127 IU/mL, but HBV DNA was negative (Sansure method). As shown in Table 1, all of HBsAg, HBcAb and HBV-PerS1 were positive at admission and the patient’s liver function was affected. However, regarding HBV, the results between immunological tests and nucleic acid detection are not always positively correlated, whichmakes clear interpretation difficult. High expression of HBsAg does not necessarily predict active *Corresponding authors: Liying Cheng andWanshanMa, Department of Clinical Laboratory, Shandong Provincial Qianfoshan Hospital, the First Hospital Affiliated with Shandong First Medical University, Jinan, Shandong, P.R. China, Phone: +86 0531 89269673, E-mail: cly_2014@163.com (L. Cheng), ws_ma@126.com (W. Ma) Hanxiang Chen, Xiaoning Zhang and Bingyang Zhang, Department of Clinical Laboratory, Shandong Provincial Qianfoshan Hospital, the First Hospital Affiliated with Shandong First Medical University, Jinan, Shandong, P.R. China Mengyuan Yu, Department of Radiotherapy, the Third Hospital Affiliated with Shandong First Medical University, Jinan, Shandong, P.R. China J Lab Med 2020; 44(6): 361–364