Detection of hepatitis B virus infection: A systematic review

Table 1 Studies describing detection of hepatitis B virus from samples other than serum or whole blood

Ref.	Year of publication	Sample used	Method used	Comments
Mendy et al[13]	2005	Dried blood spots along with serum	HBsAg detected by determine (TM) HBsAg	Comparison of DBS results with serum testing results: Sn 96%, Sp 100%
Chen et al[21]	2005	Ovarian tissue	HBsAg and HBcAg detected by immunocytochemistry and HBV DNA by PCR	Positivity rate of HBV DNA was 58.3%
van der Laan et al[17]	2007	Hepatocytes	Flow cytometric quantitation	A significant correlation was found between the percentage of infected hepatocytes and the intracellular expression level of HBsAg (R = 0.841, P < 0.001)
Goh et al[22]	2008	Cerumen and otorrhoea samples along with serum	HBsAg and HBeAg were detected by Enzyme Immunoassay and HBV DNA was detected by quantitative PCR	HBV DNA was detected in 66.7% of cerumen samples and 100% of otorrhoea samples
Chen et al[25]	2009	Parotid tissue	Serological markers by immunocytrochemistry and HBV DNA by PCR	Overall positivity rate was 54.5% to 58.3%
Nuriya et al[18]	2010	Hepatocyres	PCR - In situ hybridisation	All hepatocytes were infected with HBV in chronic liver disease
Villar et al[14]	2011	Dried blood spots	HBsAg, anti-HBc, and anti-HBs were detected by ELISA	Sn was 90.5%, 97.6%, and 78% for anti-HBc, HBsAg, anti-HBs assays, and Sp was 92.6%, 96.7%, and 97.3% for anti-HBc, HBsAg, and anti-HBs assays, respectively
Wu et al[29]	2012	Paraffin embedded intrahepatic and extrahepaticcholangiocarcinoma tissue	HBV DNA by nested PCR and HBV related antigens by immunohistochemistry method	HBV DNA and HBV antigens were detected significantly in cases of intrahepatic cholangiocarcinoma
Arora et al[24]	2012	Saliva	HBsAg was detected by ELISA	Sn 74.29% and Sp 100%
Cobo et al[27]	2012	Spent culture media and liquid nitrogen samples of oocytes and embryos	Reverse transcriptase PCR	Viral sequences were not detected in these samples from seropositive patients
Ye et al[28]	2013	Discarded test tube embryos from mothers with chronic HBV infection undergoing in vitro fertilization treatment	Single cell reverse transcriptase PCR	Detection rate was 13.2%
Eftekharian et al[23]	2013	Cerumen along with serum	HBV DNA was detected by PCR	HBV DNA was detected in 6.6% of HBsAg positive patients
Kong et al[26]	2013	Frozen renal tissue	HBsAg and HBcAg detected by immunohistochemistry	Found positive in 9 out of 500 patients of glomerulonephritis without serological evidence
Ross et al[15]	2013	Dried blood spots	HBsAG, anti HBcAg, anti HBsAg detected by Abbott Architect and HBV DNA by artus HBV LC PCR	Sensitivity was 98.6%, 97.1%, 97.5%, 93%
Alidjinou et al[16]	2014	Dried plasma spots	HBsAg and HBV DNA detected by ELISA and PCR	Sn and Sp 100% for serological markers and Sn 96%, Sp 100% for HBV DNA
Zhong et al[19]	2014	Hepatocytes	Covalently closed circular HBV DNA detected by in situ PCR	Helps to detect recurrence of HBV
Huang et al[20]	2015	Formalin fixed paraffin embedded hepatocellular carcinoma tissue	Droplet digital PCR to detect HBV copy number	Highly sensitive method

HBsAg: Hepatitis B surface antigen; DBS: Dried blood spots; Sn: Sensitivity; Sp: Specificity; HBcAg: Hepatitis B core antigen; HBV DNA: Hepatitis B virus deoxy ribonucleic acid; PCR: Polymerase chain reaction; ELISA: Enzyme linked immunosorbent assay; Anti-HBc: Antibody to HBcAg; Anti-HBs: Antibody to HBsAg.

Table 2 Studies describing different quantitative molecular methods

Ref.	Year of publication	Method of quantitation	Detection limit
Garson et al[58]	2005	FRET based real time PCR assay	Sn at 95% detection level was 24.2 IU/mL
Welzel et al[59]	2006	Novel real time PCR	Sn at 95% detection level was 56 IU/mL
Mazet-Wagner et al[60]	2006	Real time PCR assay to detect total HBV DNA and cccDNA from serum and peripheral blood mononuclear cells	27 IU/mL
McCormick et al[61]	2006	Procleix Ultrio Assay (Multiplex PCR) to detect HIV 1, HCV RNA and HBV DNA simultaneously	Sp ≥ 99.5%, Sn is > 95% with detection limit for HBV DNA of 15 IU/mL
Cai et al[62]	2008	Real time fluorogenic Loop Mediated Isothermal Amplification (RtF-LAMP)	At 95% detection level 210 copies/mL
Paraskevis et al[63]	2010	New ultrasensitive in house real time PCR assay	Sn at 95% and 50% detection level: 22.2 IU/mL and 8.4 IU/mL
Chevaliez et al[64]	2010	v2.0 of the CAP/CTM assay	Highly Sn, could even detect 6.25 IU/mL HBV DNA; Sp is 99%, Intra-assay and interassay coefficients of variation ranged from 0.21% to 2.67% and from 0.65% to 2.25%, respectively
Sun et al[65]	2011	Duplex real-time PCR assay using two sets of primers/probes and a specific armored DNA as internal control	Detection limit 29.5 IU/mL; Sp 100%
Cha et al[66]	2013	ExiStation HBV diagnostic system	9.55 IU/mL
Yang et al[67]	2014	Colorimetric PCR with DNA zyme containing probe	Broad range of linearity and high sensitivity
Kania et al[68]	2014	2 in house real time PCR targeting X (qPCR1) or S (qPCR2) genes	qPCR1: 104 IU/mL; qPCR2: 91 IU/mL

FRET: Fluoresence resonant energy transfer; PCR: Polymerase chain reaction; cccDNA: Covalently closed circular DNA; Sn: Sensitivity; Sp: Specificity; HBV DNA: Hepatitis B virus deoxy ribonucleic acid; v2.0: Version 2.0; HCV: Hepatitis C virus; CAP/CTM: CobasAmpliPrep/CobasTaqMan.

Table 3 Comparison of different methods

Ref.	Year of publication	Comparison between	Remarks
Hochberger et al[75]	2006	Automated COBAS AmpliPrep/COBAS TaqMan system (real time PCR) and Versant HBV 3.0	Good correlation between two
Juman Awadh et al[76]	2008	Versant HBV 3.0 (Bayer, branched DNA mediated assay) and Biotitre B (real time PCR variant)	Both were highly specific, though reproducibility of Versant HBV 3.0 was higher
Yang et al[77]	2009	Real Art HBV PCR Kit (Abbott, real time PCR) and Versant bDNA 3.0	Abbott's kit was more sn, detection limit 27 IU/mL
Louisirirotchanakul et al[78]	2010	fully automated ElecsysHBsAg II assay, Architect, AxSYM and Advia Centaur HBsAg assays	The later 2 tests appeared less sensitive in detecting early HBV infection
Berger et al[79]	2010	CAP/CTM assay v2.0 and v1.0	Comparable results for all 278 tested samples
Lunel-Fabiani et al[80]	2010	Access immunoassay system from Beckman coulter with Abbott AxSYM and PRISM HBsAg assays; VIDAS was used to conclude discrepant results	Sn: 100%, Sp: 99.96%
Caliendo et al[81]	2011	Abbott RealTime HBV IUO, the Roche CAP/CTM HBV test, the Roche CobasTaqMan HBV test with HighPure system, and the Qiagen artus HBV TM ASR	Limit of detection of artus 1.5 log(10) IU/mL, of other 3 tests 1.0 log(10) IU/mL
Ismail et al[82]	2011	Abbott HBV real-time PCR (Abbott PCR), artus HBV real-time PCR with QIAamp DNA blood kit purification (artus-DB), and artus HBV real-time PCR with the QIAamp DSP virus kit purification (artus-DSP)	Lower limit of detection against WHO standards were 1.43, 82 and 9 IU/mL respectively
Yeh et al[83]	2014	Abbott real time HBV (RealTime assay) and CAP/CTM HBV assays 2.0 (TaqMan assay)	Real time assay's Sn: 98.2%, Sp: 100%. Good level of agreement between the two

PCR: Polymerase chain reaction; Sn: Sensitivity; Sp: Specificity; WHO: World Health Organization; v2.0: Version 2.0; CAP/CTM: CobasAmpliPrep/CobasTaqMan; HBV: Hepatitis B virus; bDNA: Branched DNA; HBsAg: Hepatitis B surface antigen.