Blood donors screening for blood born viruses in Poland.

Blood donor screening of viral markers in Poland is based on serologic testing for anti-HCV, HBsAg, anti-HIV1/2 (chemiluminescence tests) and on nucleic acid testing (NAT) for RNA HCV, RNA HIV-1 and DNA HBV performed in minipools of 6 with real-time PCR (MPX 2.0 test on cobas s201) or with TMA in individual donations (Ultrio Plus or Ultrio Elite). Donors of plasma for anti-D and anti-HBs production are tested for parvovirus B19 DNA. Before implementation tests and equipment are evaluated at the Institute of Hematology and Transfusion Medicine (IHTM). The last 20 years witnessed a decreasing trend for HBsAg in both first time and repeat donors (1%-0.3% and 0.1%-0.02% respectively). Prevalence of anti-HCV repeat reactive results was stable and oscillated around 0.8% for first time donors and 0.2% for repeat donors. Elevated prevalence of seropositive HIV infected donors was recently observed (7.5-9 cases/100,000 donors). Since respective molecular markers implementation HCV RNA was detected on average in 1/119,235 seronegative donations, HIV RNA in 1/783,821 and HBV DNA in 1/61,047. HBV NAT yields were mostly occult hepatitis B (1/80,248); window period cases were less frequent (1/255,146). The efficiency of HBV DNA detection depends on the sensitivity of the HBV DNA screening system.

[Analysis of the prevalence of HBV markers relevant to the risk of reactivation of infection in patients with hematologic diseases]. / Analiza wystepowania markerów wirusa HBV istotnych dla ryzyka reaktywacji zakazenia u osób z chorobami hematologicznymi.

The study assessed the incidence of HBV markers (HBsAg, anti-HBc, anti-HBs) important for determination of the risk of reactivation of infection, with particular interest of occult infection (presence of HBV DNA in the absence of HBsAg) in patients treated at the Institute of Hematology and Transfusion Medicine. Anti-HBc frequency was correlated with the age and sex of patients. HBsAg was detected in 16/468 examined patients, 98/468 (21%) were anti-HBc positive. HBV DNA was detected in 41/98 anti-HBc positives; in 13 simultaneously with HBsAg. 28 patients had occult HBV infection (HBV DNA+/HBsAg). Antibody to HBsAg was detected in 163/430 (38%) patients, 81 out of them on protective level (> 100 IU/l). It was shown that occult HBV infection occurs in approximately 6% of patients. In most of them the protective levels of anti-HBs are detected.

Unexpected Course of [2+3] Cycloaddition of 2-nitropropene to (Z)-C,N-diphenylnitrone.

The [2+3] cycloaddition of 2-nitropropene to (Z)-C,N-diphenylnitrone leads to 3,4-trans-2,3-diphenyl-4-nitro-4-methyl- and 3,5-trans-2,3-diphenyl-5-nitro-5-methylisoxazolidines as primary reaction products. This, however, is not the only pathway of 2-nitropropene conversion. In the reaction conditions, the nitroalkene also undergoes isomerisation and the resulting trans- and cis-1-nitropropenes yield respective stereoisomeric 2,3-diphenyl-4-nitro-5-methylisoxazolidines in the reaction with (Z)-C,N-diphenylnitrone.

[Prevalence of HCV, HBV and HIV infections among severe Polish haemophiliacs]. / Czestosc zakazen HCV, HBV i HIV u chorych na ciezka hemofilie w Polsce: porównanie chorych urodzonych przed i po 1991 roku.

UNLABELLED: In the past, haemophilia replacement therapy was based on fresh frozen plasma, cryoprecipitate and blood transfusions--i.e. preparates not subjected to any viral inactivation methods. The aim of the present study was to assess the prevalence of HCV, HBV and HIV infections among Polish haemophiliacs. MATERIALS AND METHODS: Severe haemophilia patients were divided into two age groups according to the type of replacement therapy used in the past: group 1--172 patients born 1935-1990, group 2--41 patients born 1991-2002. The following viral markers were tested: anti-HCV, RNA HCV (if needed--virus genotype), HBsAg and anti-HIV. In 75 patients / group 1 and 41 / group 2, anti-HBc presence was determined. RESULTS: In group 1, 95% of patients were anti-HCV positive and in 77.3% RNA HCV was detected. The most prevalent was lb genotype (58.6%) followed by 3a (27%). HBsAg was found in 8.7% cases of group 1. Anti-HCV antibodies were detected in 51 of 75 (68%) patients of this group. One patient was anti-HIV positive. In only one patient of group 2 (2.4%) viral markers were detected (anti-HCV and RNA HCV). CONCLUSION: Nowadays, the risk of HCV, HBV and HIV infection in Polish severe haemophiliacs is very low.

Characterization of HBV DNA+/HBsAg- blood donors in Poland identified by triplex NAT.

Nucleic acid testing (NAT) for hepatitis B virus (HBV) has been performed in Poland since 2005 on samples seronegative for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV), and anti-human immunodeficiency virus (anti-HIV). Tools included 24-donation pool testing (PT) using Cobas Amplicor or in individual donations (ID) by Procleix Ultrio. Seven of 761,666 (1:108,800) and 21/250,191 (1:11,900) HBV DNA-positive donations were identified and confirmed by alternative methods. HBV DNA load ranged between 11.6 and 4.6 x 10(4) IU/mL in 11 samples and could not be quantified in 17 samples. HBV genotypes A (56%) and D (4%) were found. The analysis of combined results from index, follow-up, and look-back samples identified four groups: (1) Two cases tested HBsAg positive with alternative, more sensitive, assays; (2) Four cases were in the pre-seroconversion window period; (3) Eight cases had a fluctuating pattern of HBV DNA and anti-HBs detection (recovered infection); and (4) twelve cases carried anti-HBc without anti-HBs, which might correspond to either chronic or recovered "occult" HBV infection. One donor with no HBV markers in the follow-up was excluded, and another was in the window period preceding anti-HBs. HBV NAT identified more confirmed positive donors than HCV or HIV NAT, and 1:250,000 could not be detected by anti-HBc screening. Serological and molecular studies on follow-up and look-back samples are important to classify donors. In conclusion, further studies are needed to determine whether the considerably higher yield of HBV DNA detection obtained with individual donation screening improves blood safety compared with anti-HBc screening.

[GBV-C/HGV and TTV infection markers in Polish blood donors and haemophilia patients]. / Markery zakazenia wirusami GBV-C/HGV i TTV u chorych na hemofilif, oraz u dawców krwi w Polsce.

Viruses GBV-C/HGV and TTV were identified in patients with hepatitis of unknown etiology. Aim of our study was to assess the frequency of infection markers of these viruses in blood donors and haemophilia patients treated with virucidaly activated and non inactivated blood products. Material and methods. TTV DNA (by PCR using primers to coding ORFI and non-coding region NC) and GBV-C/HGV (RNA by RT-PCR and anti-E2 by EIA) were tested in blood donors (200 for TTV and 219 for GBV-C/HGV), 122 haemophilia patients treated in the past with non inactivated blood products and in 20 haemophilia children treated exclusively with inactivated clotting preparations. Results RNA GBV-C/HGV were identified in 3,2%; 23,7% and in 0%, respectively blood donors, adult and children haemophilia patients. Antibody anti-E2 were found in 23,6%; 37% i 25% of studied groups respectively. DNA TTV was detected most frequently by NC than ORF1 primers: in 78% vs.10% of blood donors, 100% vs 43,5% of adult haemophilia patients and in 95% vs. 15% young haemophiliacs. Conclusions Haemophilia patients were at risk of GBV-C/HGV and TTV infection. Following implementation of viral inactivation methods in the process of clotting factor concentrates production, the risk for GBV-C/HGV transmission was significantly reduced and in less extant for TTV.

Hepatitis C core antigen in Polish blood donors.

BACKGROUND: The goal of this study was to evaluate the feasibility of adopting the HCV core antigen ELISA (HCVcAg) for routine screening of Polish blood donors. STUDY DESIGN AND METHODS: A total of 133,279 donor samples were tested by ORTHO HCVcAg. All repeatedly reactive (RR) samples were tested by neutralization test for confirmation, RIBA HCV for anti-HCV, and by Cobas Amplicore for HCV RNA. All donations were tested for ALT level. RESULTS: The HCVcAg test specificity was 99.94 percent. In total, 1499 donations (1.12%) were initially reactive and 124 (0.09%) were RR. Antibodies to HCV were found in 22 out of 124 donors and HCV RNA was detected in 19 out of 22. In 10 out of the 19 HCV-RNA-positive donors, the HCVcAg neutralization test was positive. Among the 102 HCVcAg RR/anti-HCV-negative donors, there were 6 neutralization-test-positive individuals, and all were HCV RNA positive. Elevated ALT level was observed in one of them. During the follow-up studies of three HCVcAg RR/HCV-RNA-positive donors, seroconvertion was observed 5 to 7 weeks after the initial HCVcAg-positive result. In all, HCVcAg results became negative once antibodies to HCV were detected. CONCLUSION: The HCVcAg test proved to be feasible for routine screening in the Polish Blood Transfusion Service. Six HCVcAg RR/anti-HCV-negative donors were identified. The calculated residual risk in this study of donors in the preseroconversion window was 45 per million. Mandatory testing of every blood and plasma donation for HCVcAg or HCV RNA was recommended as of January 2, 2002.