The utility of a gross dissection anatomical model for simulation-based learning in pathology.

INTRODUCTION: The examination of morphological alterations in tissues is fundamental in Pathology. Traditional training in gross dissection has several limitations, including the risk of transmissible diseases, formaldehyde exposure and limited specimen availability. We describe a teaching method using anatomical simulators. METHODS: Liquid silicone-based artisan neoplastic anatomical models were used in conjunction with clinical scenarios. Eighty-five medical students participated in a gross dissection experience and were asked to complete a feedback questionnaire. Additionally, a workshop was organized for students to compare three different teaching methods. The first one used still images (Group1-G1), the second a video explanation (Group2-G2), and the third directly observed a pathologist while grossing (Group3-G3). RESULTS: The knowledge acquisition questionnaire showed an average value of 4.4 out of 5 (1-5) (range 3.4-4.7, σ0.89). The categories 'knowledge of resection margins' and 'macroscopic diagnosis' received the highest values (4.8, σ0.11 and 4.7, σ0.32, respectively), followed by 'understanding of handling and gross examination of the surgical specimen' (4.5, σ0.49), 'prognosis' (4.3, σ0.67) and 'understanding of a tumor resection' (3.9, σ0.96) (p<0.05). Regarding teaching methods, G3 spent less time than G2 and G1 with mean times of 15'39″ (σ2'12″), 16'50″ (σ3'45″), and 17'52″ (σ2'12″), respectively (p<0.05). Gross dissection marks (0-5) showed statistically significant differences (p<0.05). G2 obtained better results (3.7;σ0.54) than G3 (3.4;σ0.94) or G1 (3.1;σ0.8). CONCLUSIONS: This preliminary study demonstrates that it is possible to implement a gross dissection simulation module at medical school and thus enable the acquisition of skills in a secure environment.

Mid-stream vs. first-voided urine collection by using automated analyzers for particle examination in healthy subjects: an Italian multicenter study.

BACKGROUND: In analogy with other areas of laboratory diagnostics, the pre-analytical phase is the leading source of variability also in urinalysis. We carried out a multicentric study for comparing results obtained from first-voided and mid-stream urine samples. METHODS: Each of the six hospital-based clinical laboratories participating to this study recruited 50 healthy subjects among laboratory staff and/or their relatives. Two consecutive samples of the first morning micturition were collected by vacuum system, the first from the first-void and the second from the mid-stream. Routine urinalysis was performed using dip-stick automated analyzers for chemical examination and automated analyzers for formed particle examination (Sysmex UF-100, Sysmex UF-1000i and Iris iQ-200). RESULTS: Counts of epithelial cells (EC), erythrocytes (ERY) and leukocytes (LEU) but not for cylinders (CAS) were significantly higher in the first-voided samples. A significantly higher count of EC, ERY and LEU was also observed between females and males in first-voided samples, whereas no significant difference could be found in mid-stream samples. Health related analyzer specific upper reference limits (URL) were CAS≤1, EC≤5, ERY≤19, Leu≤13 for UF-100; CAS≤1, EC≤4, ERY≤15, Leu≤11 for UF-1000i; CAS≤1, EC≤4, ERY≤18, Leu≤10 for iQ200. The overall prevalence of subjects with cellular elements count exceeding URL was also higher in first-voided than in mid-stream samples. CONCLUSIONS: Mid-stream urine was confirmed as the most appropriate sample, since the presence of contaminating elements, such as bacteria, analytes and formed particles are minimized.

Urine particle evaluation: a comparison between the UF-1000i and quantitative microscopy.

BACKGROUND: The study of urine particles plays a key role in the diagnosis of kidney diseases. In this study, the authors evaluated the correlation between the UF-1000i and quantitative manual microscopy. METHODS: A total of 214 untreated urine samples were studied using the Sysmex UF-1000i and compared with results obtained from quantitative manual microscopy using the Fuchs-Rosenthal counting chamber. RESULTS: Using Pearson statistics, we observed satisfactory correlation between the UF-1000i and quantitative microscopy: for red blood cells (RBCs) r was 0.98, for white blood cells (WBCs) r was 1.00, for epithelial cells (EC) r was 0.96, and for casts r was 0.69. Using linear regression statistics, we also observed satisfactory correlation between the UF-1000i and quantitative microscopy: for RBCs R(2) was 0.95, for WBCs R(2) was 0.99, for EC R(2) was 0.92, and for casts R(2) was 0.48. CONCLUSIONS: In our experience, automated urine particle analysis performed using the Sysmex UF-1000i analyzer is sufficiently precise and improves the workflow in a routine laboratory. Precision was satisfactory and concordance with the reference method is good for RBC, WBC and EC; for casts microscopic observation is required for flagged samples to discriminate hyaline from pathologic casts.

[Thrombophilia risk factors evaluation in a group of Italian patients with deep venous thrombosis]. / Valutazione del rischio trombofilico in una coorte di pazienti con trombosi venosa profonda.

UNLABELLED: Venous thrombosis usually results from coexistence of multiple genetic and acquired risk factors with a trigger condition. In this study the authors report their experience in a cohort of Italian patients with previous venous thrombosis. MATERIAL AND METHODS: We considered 292 consecutive patients. Each patient was studied by using a panel of functional and genetic tests to detect some of the most relevant thrombophilia risk factors. RESULTS: The single most frequent thrombophilia risk factor was activated C protein resistance due to FV Leiden. Tests for anti phospholipids auto antibodies showed reactivity in 62 subjects. CONCLUSION: The great majority (80%) of patients showed almost one thrombophilia risk factor. Presence of multiple risk factors was demonstrated in 128 (44%) patients.

[PHYSICAL, CHEMICAL AND MORPHOLOGICAL URINE EXAMINATION: RECOMMENDATIONS FOR THE POST ANALYTICAL PHASE FROM THE INTERDISCIPLINARY URINALYSIS GROUP (GIAU)].

With these recommendations the Interdisciplinary Urinalysis Group (GIAU) aims to stimulate the following aspects : improvement and standardization of the post analytical approach to physical, chemical and morphological urine examination (ECMU); emphasize the value added to ECMU by selection of clinically significant parameters, indication of analytical methods, of units of measurement, of reference values; improvement of interpretation of dip stick urinalysis with particular regard to the reconsideration of the diagnostic significance of the evaluated parameters together with an increasing awareness of the limits of sensitivity and specificity of this analytical method. Accompanied by the skills to propose and carry out in-depth investigations with analytical methods that are more sensitive and specific;increase the awareness of the importance of professional skills in the field of urinary morphology and their relationships with the clinicians. through the introduction, in the report, of descriptive and interpretative comments depending on the type of request, the complexity of the laboratory, the competence of the pathologist;implement a policy of evaluation of the analytical quality by using, in addition to traditional internal and external controls, a program for the evaluation of morphological competence. The hope is to revalue the enormous potential diagnostic of ECMU, implementing a urinalysis on personalized diagnostic needs that each patient brings with it.

Clinical evaluation of a functional prothrombin time-based assay for identification of factor V Leiden carriers in a group of Italian patients with venous thrombosis.

The efficiency of a new prothrombin-based activated protein C (APC) resistance test to detect factor V Leiden (FVL) was clinically evaluated in 150 Italian patients with deep venous thrombosis. Patient samples are diluted in factor-V-deficient plasma, an APC-containing reagent, and specific factor V activator; after incubation, clotting is initiated by addition of activated-factor-FV-dependent prothrombin activator. Two prothrombin time determinations were performed under identical assay conditions except that no APC was added to one. A ratio over 4.2 for normal individuals and under 2.0 for FVL patients is expected: between 1.3 and 1.9 for FVL heterozygotes, and between 1.0 and 1.1 for FVL homozygotes. Using a predefined cut-off ratio of 2.0, a specificity and a sensitivity of 1.00 for detection of FVL mutation were found. With a cut-off ratio of 1.1, a specificity of 0.98 and a sensitivity of 1.00 were found for discrimination between FVL heterozygous (n = 60) and homozygous (n = 6). No interferences by heparins, oral contraceptives, oral anticoagulant therapy, protein C, protein S, D-dimer, homocysteine, MTHFR mutations and antiphospholipid autoantibodies were detected. In our experience, this new prothrombin time-based APC resistance assay provides improved discrimination between normal individuals and FVL carriers compared with the classical methods. Moreover, this new assay allows good discrimination between homozygous and heterozygous FVL carriers. In the authors' experience this prothrombin time-based method was not influenced by many factors compared with the classical activated partial thromboplastin time-based method.

Effect of dabigatran on a prothrombinase-based assay for detecting activated protein C resistance: an ex vivo and in vitro study in normal subjects and factor V Leiden carriers.

BACKGROUND: The aim of this study was to evaluate ex vivo and in vitro interference of a direct factor IIa inhibitor, dabigatran, on a prothrombinase-based assay to detect activated protein C resistance. MATERIALS AND METHODS: An ex vivo study was performed in six heterozygous factor V Leiden carriers and 12 normal subjects without the factor V Leiden mutation who were treated with dabigatran. An in vitro study was also performed considering 12 plasma samples (six from normal subjects and six from heterozygous factor V Leiden carriers) spiked with dabigatran. The dabigatran concentration was evaluated using a diluted thrombin time assay, activated protein C resistance was evaluated using a prothrombinase-based assay. RESULTS: In both the ex vivo and in vitro studies dabigatran interfered significantly with activated protein C resistance ratios observed in normal subjects and in factor V Leiden heterozygous carriers. DISCUSSION: The results reported in this paper seem to confirm that dabigatran is able to interfere with the Penthafarm prothrombinase-based assay used to study activated protein C resistance, significantly increasing observed ratios. This effect appears to be present already at low concentrations of dabigatran (6 ng/mL) and affects both normal subjects and heterozygous carriers of factor V Leiden. In this group of patients, dabigatran, at concentrations in the therapeutic range (100-200 ng/mL), could markedly increase the activated protein C resistance ratio, bringing it up to within the reference range for normal subjects, thus potentially leading to misclassification of patients.

Estimated glomerular filtration rate calculated using different creatinine and cystatin based formulas in prediction of trough plasma Dabigtran concentration.

BACKGROUND: In this paper we evaluated the relationship between trough DAB concentration with eGFR calculated using CRE and CYS based formulas. MATERIALS AND METHODS: We considered 100 patients. eGFR was calculated using CKD-EPIcreat, CG, MDRD, CKD-EPIcys and CKD-EPIcombined equations. DAB dosage was selected on the basis of CKD-EPIcreat and relationship between trough DAB concentration and eGFRs was evaluated. RESULTS: Trough DAB concentration roughly correlates with eGFR calculated using various formulas. CKD-EPIcreat eGFR was higher than CKD-EPIcys. In patients receiving a DAB dosage considered adequate using CKD-EPIcreat eGFR but considered excessive using CKD-EPIcys, we observed higher DAB trough concentration and an increased prevalence of subjects with drug concentration >200ng/mL. CONCLUSION: These results suggest that eGFR alone was unable to fully explain trough DAB plasma concentration. Therefore a drug's prescription schedule based on eGFR only should be inadequate. We observed a better correlation between trough DAB concentration and CKD-EPIcys rather than CKD-EPIcreat eGFR. Thus, in patients chronically treated with DAB for thromboprophylaxis in nonvalvular atrial fibrillation evaluation of eGFR using a cystatin base formula should be considered.

[Physical, chemical and morphological urine examination guidelines for the Analytical Phase from the Intersociety Urinalysis Group]. / Esame fisico, chimico e morfologico delle urine proposta di linee guida per la fase analitica del Gruppo Intersocietario Analisi delle Urine (GIAU).

With these guidelines the Intersociety Urinalysis Group (GIAU) aims to stimulate the following aspects: Improvement and standardization of the analytical approach to physical, chemical and morphological urine examination (ECMU). Improvement of the chemical analysis of urine with particular regard to the reconsideration of the diagnostic significance of the parameters that are traditionally evaluated in dipstick analysis together with an increasing awareness of the limits of sensitivity and specificity of this analytical method. Increase the awareness of the importance of professional skills in the field of urinary morphology and the relationship with the clinicians. Implement a policy of evaluation of the analytical quality by using, in addition to traditional internal and external controls, a program for the evaluation of morphological competence. Stimulate the diagnostics industry to focus research efforts and development methodology and instrumental catering on the needs of clinical diagnosis. The hope is to revalue the enormous diagnostic potential of 'ECMU, implementing a urinalysis on personalized diagnostic needs for each patient. Emphasize the value added to ECMU by automated analyzers for the study of the morphology of the corpuscular fraction urine. The hope is to revalue the enormous potential diagnostic of 'ECMU, implementing a urinalysis on personalized diagnostic needs that each patient brings with it.

Does flow cytometry have a role in preliminary differentiation between urinary tract infections sustained by gram positive and gram negative bacteria? An Italian polycentric study.

BACKGROUND: Urine culture is the most frequently requested test for a Microbiology Lab. A reliable screening tool would be of paramount importance both to clinicians and laboratorians, provided that it could get fast and accurate negative results in order to rule-out urinary tract infection (UTI). MATERIALS AND METHODS: We evaluated 1907 consecutive urine samples from outpatients. Culture was performed on chromogenic agar with 1µL loop, using 10(5)CFU/mL as a limit of positive growth. Using Sysmex Uf-1000i analyzer we evaluated bacteria forward scatter (B_FSC) and fluorescent light scatter (B_FLH) in a preliminary discrimination step for UTI caused by Gram+ or Gram- bacteria. RESULTS: We got 512 positive samples. A mono-microbial infection was observed in 490 samples; two bacterial strains were isolated in 22 samples, so 534 bacterial strains were found: 392 Gram-, 133 Gram+ and 9 yeasts. Comparing Gram+ and Gram- bacteria we observed a statistically significant difference for B_FSC but not for B_FLH. In this application experimental cut-off value for B_FSC was 25ch. Using this cut-off to perform a presumptive identification of UTI sustained by Gram-+ bacteria, we observed a SE 0.68, SP 0.84. CONCLUSION: Our data although preliminary suggest that B_FSC could be useful in presumptive exclusion of UTI caused by Gram-positive bacteria.

Postpartum thrombotic complication in a patient with paroxysmal nocturnal hemoglobinuria.

Paroxysmal nocturnal hemoglobinuria (PNH) is a clonal nonneoplastic hematopoietic stem cell disease characterized by an acquired mutation of the PIG-A gene with reduction or absence of CD55 and CD59. The absence of these proteins renders PNH erythrocytes susceptible to complement-mediated hemolysis. We report the case of a PNH patient before and during pregnancy until delivery. We observed and treated some postpartum thrombotic complications. Eculizumab should be used with caution in pregnancy. There are several reports supporting its use in these patients. This case should be considered paradigmatic of a series of clinical situations that may occur in the course of a pregnancy in patients with PNH: increased need for transfusion, need to increase the dose of Eculizumab, and insurgence of fetal sufferance. Moreover, after delivery, the patient, despite adequate prophylaxis with low-molecular-weight heparins, presented severe complications: development of pleural and peritoneal effusion, pulmonary embolism, bilateral upper limbs thrombophlebitis, and a possible abdominal angina with a transient paralytic ileus. All these complications were overcome and now the baby is healthy and the mother has returned to the usual therapeutic regimen.

Significance of anti-HBc only in blood donors: a serological and virological study after hepatitis B vaccination.

BACKGROUND: Blood donors positive only for anti-HBc may have a resolved hepatitis B virus (HBV) infection, low grade chronic infection or infection with variant strains of HBV. We aimed to assess the significance of this serological pattern after hepatitis B vaccination in such cases. MATERIALS AND METHODS: Twenty-four anti-HBc only blood donors were vaccinated with the Engerix HBV vaccine and a serological and virological evaluation was performed before HBV vaccination and 7-10 days after each dose. Subjects were classified as non-responders if their anti-HBs levels stayed below 10 IU/L after full vaccination, while the response was considered secondary (anamnestic) if anti-HBs levels rose over 10 IU/L after the first vaccine dose, and primary if anti-HBs levels rose over 10 IU/L only after the second or third vaccine dose. RESULTS: Of the 21 fully evaluable donors, six had no response, eight showed a primary response and seven had an anamnestic response. One non-responder had transient positivity for HBV-DNA at low levels (12 IU/mL) with persistent negativity for HBsAg. DISCUSSION: Anti-HBc-only positive blood donors are a heterogeneous population including HBV naïve subjects with a likely false-positive anti-HBc reactivity, subjects with a resolved HBV infection, and subjects with persistent low-level HBV replication. The analysis of the anti-HBs response after a dose of HBV vaccine may help to distinguish among the different causes of the isolated anti-HBc positivity, thereby enabling proper counselling and potential readmission to blood donation.

Gender's equality in evaluation of urine particles: Results of a multicenter study of the Italian Urinalysis Group.

OBJECTIVE: We performed a multicenter study to calculate the upper reference limits (URL) for urine particle quantification in mid-stream samples by using automated urine analyzers. DESIGN & METHODS: Two laboratories tested 283 subjects using a Sysmex UF-100, two other laboratories tested 313 subjects using Sysmex UF-1000i, whereas two other laboratories tested 267 subjects using Iris IQ®200. RESULTS: The URLs of UF-100 in females and males were 7.8/µL and 6.7/µL for epithelial cells (EC), 11.1/µL and 9.9/µL for red blood cells (RBC), 10.2/µL and 9.7/µL for white blood cells (WBC), and 0.85/µL and 0.87/µL for cylinders (CAST). The URLs of UF-1000i in females and males were 7.6/µL and 7.1/µL for EC, 12.2/µL and 11.1/µL for RBC, 11.9/µL and 11.7/µL for WBC, and 0.88/µL and 0.86/µL for CAST. The URLs of Iris IQ®200 in females and males were 7.8/µL and 6.6/µL for EC, 12.4/µL and 10.1/µL for RBC, 10.9/µL and 9.9/µL for WBC, and 1.1/µL and 1.0/µL for CAST. CONCLUSION: The URLs obtained in this study were comparable to the lowest values previously reported in the literature. Moreover, no gender-related difference was observed, and analyzer-specific upper reference limits were very similar.

Pediatric reference values for urine particle quantification by using automated flow cytometer: results of a multicenter study of Italian urinalysis group.

OBJECTIVES: The purpose of this Italian multicenter study was to define pediatric upper reference values for urine particle quantification by using automated flow cytometry. DESIGN AND METHODS: Four hospital-based clinical laboratories participated in this multicenter investigation, which included a total study population of 161 Italian children aged from 1 to 12years. Two laboratories used Sysmex UF-100 and analyzed 86 children, whereas the other two used Sysmex UF-1000i and analyzed 75 subjects. Particle quantification included the analysis of white blood cells (WBC), red blood cells (RBC), squamous epithelial cells (EC), transitional epithelial cells (TC), casts (CAST) and bacteria (BACT). RESULTS: The upper reference values in subjects tested with the Sysmex UF-100 were 9.7WBC/µL, 10.1RBC/µL, 7.5EC/µL, 2.5TC/µL, 0.7CAST/µL and 3090BACT/µL, whereas the upper reference values in subjects tested with the Sysmex UF-1000i were 10.5WBC/µL, 8.3RBC/µL, 7.2EC/µL, 2.9TC/µL, 0.7CAST/µL and 48BACT/µL. No statistically significant differences between genders were found in the value distribution of any of the parameters tested. Similarly, no statistically significant differences were observed between the two urine analyzers, except for BACT. CONCLUSIONS: Automated analysis of urine particles appears a suitable means to optimize the workflow of routine urinalysis of children specimens. The upper reference limits for pediatric subjects obtained in this study were comparable to those previously reported in the literature, with no significant differences between genders and analyzers.

Case-control time course study of urinary leukocyte and bacterial counts in patients with acute urinary tract infections responsive and not responsive to antibacterial therapy.

BACKGROUND: Urinary tract infections (UTIs) are usually diagnosed by urine particle detection and urine bacterial culturing, but an empiric treatment is often prescribed before the microbiological results are known. The aim of this study was to assess whether quantification of bacteriuria and leukocyturia could predict the outcome of empiric antibiotic therapy. METHODS: By using a Sysmex UF-100 analyser, we performed a case-control study in which bact channel counts (BCCs) and leukocyte (WBC) quantification were compared in urine samples obtained from responsive patients (RPs) and non-responsive patients (NRPs) at diagnosis (t0) and 24h (t24) after the start of empiric antibacterial treatment. RESULTS: BCCs decreased significantly from t0 (median: 30,000 E6/L) to t24 (median: 700 E6/L; p<0.001) in RPs but not in NRPs (median at t0 31,000 E6/L, and 23,000 E6/L at t24; p>0.05). Similarly, WBC counts were reduced in RPs at t24 relative to the initial values (p<0.001) but not in NRPs. A reduction of 70% in the BCC count or 60% in the WBC count of paired samples was useful for discriminating between RPs and NRPs. CONCLUSIONS: Our results suggest that quantitative time course analysis of urine BCCs and WBCs can be used to predict the success of first-line empiric treatment for UTIs.

Performance of automated platelet quantification using different analysers in comparison with an immunological reference method in thrombocytopenic patients.

BACKGROUND: Rapidly available and accurate platelet counts play an important role in the evaluation of haemorrhagic status and in assessing the need for platelet transfusions. We, therefore, evaluated platelet counting performance of haematology analysers using optical, impedance and immunological methods in thrombocytopenic patients. MATERIALS AND METHODS: We considered 99 patients with a platelet (plt) count under 50 x 10(9) plt/L. We compared the platelet counts obtained using ADVIA 2120 (optical method), Cell-Dyn Sapphire (optical, impedance and immunological methods with CD61) and a reference, double staining (CD41+CD61) immunological method. RESULTS: The platelet counts of all the considered methods showed good correlation with those of the reference method, despite an overestimation in platelet quantification. The degree of inaccuracy was greater for platelet counts under 20 x10(9) plt/L. CONCLUSIONS: Clinicians who use platelet thresholds below 20 x10(9) plt/L for making clinical decisions must be aware of the limitations in precision and accuracy of cell counters at this level of platelet count. Inaccurate counts of low platelet numbers could create problems if attempts are made to reduce the threshold below 20 x 10(9) plt/L.

Cutoff values for bacteria and leukocytes for urine flow cytometer Sysmex UF-1000i in urinary tract infections.

Because urinary tract infections (UTIs) are a quite common disease, the gold standard for diagnosing UTIs is still bacterial culture, although a large percentage of samples are negative: unnecessary cultures can be reduced by means of an effective screening test. The analytic performance of a new urine cytometer, the UF-1000i, has been tested on 1463 urine samples submitted to our laboratory for culture. Bacteria and leukocyte counts have been compared by means of the UF-1000i with colony-forming unit (CFU) quantification on citrate lactose electrolytes deficient agar to assess the best cutoff values. By using quantitative cultures and considering as positive a sample with 10 x 10(5) CFU/mL, 546 positive samples (37%) were observed. If compared with 10 x 10(5) CFU/mL, the cutoff values obtained were 125 bacteria/microL and 40 leukocytes/ microL, respectively. Analytic parameters such as sensitivity, specificity, positive predictive value, negative predictive value, and correctly classified incidence were satisfactory. Based on the results obtained in this study, when using the UF-1000i analyzer for a screening test for UTI, a cutoff value of 40 white blood cells/microL should be adopted. The cutoff value for bacteria should be 125/microL for those clinical conditions in which 10 x 10(5) CFU/mL indicates a positivity.

What is the role of serology for the study of chronic hepatitis B virus infection in the age of molecular biology?

To assess quantitative serology in chronic hepatitis B virus (HBV) infection, testing by novel immunoassays has been carried out on 202 specimens from untreated patients and in 83 samples from 10 patients with chronic hepatitis B treated with lamivudine. Serum samples were assayed for quantitative HBsAg, in comparison with quantitative HBV-DNA, and for anti-HBc IgM and the avidity index (AI) of total anti-HBc antibodies. The AI was high (mean: 0.93 +/- 0.19) in all groups, confirming the consistency of this procedure in chronic HBV infections. A low-level positivity (2-28 Paul-Ehrlich units/ml) for IgM anti-HBc was detectable both in HBeAg-positive and in HBeAg-negative untreated chronic hepatitis cases (mean S/CO values by the Abbott Architect assay: 0.51 +/- 0.12 and 0.48 +/- 0.10, respectively; correlation between assays: r = 0.685), while treated patients (mean: 0.20 +/- 0.15) and inactive carriers (mean: 0.17 +/- 0.21), were generally negative for IgM. The levels of HBsAg (IU/ml) showed a weak correlation with HBV-DNA (IU/ml). A difference in HBsAg levels was found between inactive carriers (1,935 +/- 2,887 IU/ml) and chronic hepatitis B cases, either treated (5,199 +/- 9,259 IU/ml) or untreated (14,596 +/- 15,227 IU/ml). Pre-treatment levels of HBsAg in patients undergoing lamivudine treatment were correlated with a sustained response to therapy over 13-33 months (mean: 27.3) of follow-up: mean HBsAg values were 1,576 + 1,487 IU/ml in five responders and 6,063 + 5,142 in five nonresponders or breakthrough responders (P < 0.05). The availability of standardized quantitative immunoassays for HBsAg and anti-HBc IgM may be considered in addition to quantitative HBV-DNA in the staging and monitoring of chronic HBV infection.