Scientific Collaborations

Scientific collaborations


Collaborations with the Sano-Hep Association, ROMANIA

SANO-HEP is the first association of patients registered with liver diseases in Romania and a founding member of ELPA – European Liver Patients Association. SANO-HEP was established in Brasov in 2001, as a result of the need to create an adequate organizational framework among the approximately 1 million Romanian patients registered with liver and related diseases. The existence of the association is the consequence of the need of patients with such conditions to know the treatment methods and procedures used in the therapy of diseases of this type. SANO-HEP actively collaborates with the Ministry of Health and the National Health Insurance House to unconditionally support patients with liver diseases in Romania and prevent liver diseases.

Association of Transplanted Patients from Romania

At the Introduction of Molecular Biology techniques in Transplant Immunology, instead of the current text, insert the text below and only the conclusions remain.Next-generation HLA DNA sequencing NGS versus Oxford Nanopore technology to improve donor-recipient immunogenetics (2023) Donor sequencing through Nanopore technology will improve and make it possible to obtain results in only 4 hours compared to NGS where results can be given in 6 days.

HLA DNA sequencing versus HLA SSO/SSP genotyping for improving donor-recipient immunogenetics (2004)

To improve the donor-recipient HLA match, 3 molecular biology methods were evaluated in terms of HLA genotyping for kidney donors and peripheral hematopoietic stem cell donors. We were interested in the impact of HLA matching at the allele level for clinical diagnosis and for the prognosis of kidney and bone marrow allograft survival. The comparison of HLA DNA direct sequencing – SSP/SSO HLA genotyping revealed a matching performance of HLA alleles with different degrees of expression for direct sequencing between donors and recipients.


  • Direct HLA sequencing resolved all SSO/SSP allelic ambiguities in 48% of cases (donor-recipients)
  • In 4% of cases, direct HLA sequencing deciphered new alleles.


Preliminary data demonstrate that optimal HLA matching still has a lot to offer in terms of reducing the frequency of acute rejection and has an indisputable impact on the long-term survival of renal and bone marrow allografts. Through direct sequencing, there is a chance of discovering new HLA alleles in both the donor and the recipient. A more accurate HLA match allows avoiding increased doses of immunosuppressive agents and their adverse effects in the post-transplant period, thus improving the patient’s quality of life. Direct HLA sequencing: identifies null alleles, identifies insertions and deletions, resolves heterozygous ambiguities, identifies weakly expressed alleles, detects the soluble form of alleles.

The preliminary conclusions of HBV genotyping in Romania are: HBV genotype A is associated with inactive, asymptomatic carrier status and is present in double HBV-VHD infection.

HBV genotype D is associated with active viral infection and evolution towards hepatocellular carcinoma; genotype D is associated with the development of hepatocellular carcinoma.

Currently, several mutations are known that occur at the level of nucleotide sequences in the HBV genome, during the infection, and which have important clinical and virological consequences.
Thus, the mutation in the S gene that includes the substitution of glycine with arginine at codon position 145 (G145R) was frequently encountered in newborns from HBsAg positive mothers, in whom HBV infection developed despite vaccination at birth. Also, this mutation was detected in liver transplant patients who had recurrent HBV infection even though HIBg immunoglobulin was administered to them. The “core promoter” and “precore” variants produce HBV virions that do not secrete HBeAg.
The most frequent “precore promoter” mutation has a dual change A1762T and G1764A that decreases “precore” mRNA and thus HBeAg secretion.

These variants are found in particular in patients who do not have serum HBeAg present, having HBV genotypes B, C, and D. The G1896A mutation, premature stop codon, should also be mentioned.

In severe cases of acute and chronic hepatitis, “precore” HBV mutants have been described.
Mutations in the polymerase gene have been identified in patients who are on antiviral therapy and who develop antiviral resistance. The best characterized mutants in the polymerase gene were identified during lamivudine therapy.

The mutations M552V, M552I, L528M, (YMDD) occur in the catalytic domain of the polymerase that confers resistance to lamivudine and related nucleoside analogues.

Preliminary sequencing data of circulating strains of HBV in Romania were relevant by our working group and colleagues from Great Britain (I. Constantinescu, TJ Harrison, R. Ling – unpublished data, 2000).

The patients with HBV infection selected for sequencing have different forms of the disease, either aggressive or mild biochemically and histologically, our goal being to find a possible correlation between possible HBV mutants in the “core promoter” region and clinical and paraclinical aspects.

The preliminary data showed in these cases stop codon type mutations: 28 Tryptophan -> stop codon; 131 Arginine -> Lysine (aac->aaa); 145 Glycine-> Arginine (gga->aga).

This study is ongoing and the final results will provide new aspects related to mutations in the “core promoter” region and the clinical and histopathological evolution of chronic infection induced by HBV.

Molecular virology studies of HBV provide a unique understanding of the virus-host relationship with an important impact on the evolution of hepatitis infection and prognosis, but especially on the selection of patients for antiviral therapy.

In conclusion, without a molecular virology diagnosis we cannot successfully understand and treat a chronic HBV infection.

HBV carries out its DNA genome replication process via a reverse transcription process that has gaps in the reading activity, thus reaching the quasi-species form of HBV.
HBV genotypes A and D have different replication efficiencies in cell cultures and in vivo these characteristics are preserved.

Frequently, the mutant forms (especially those in the “precore”, “core promoter” region) come to predominate over time in the wild virus strain.

For viral dominance, both viral intrapopulation and intra-host factors must be taken into consideration. Mutants can develop a selection advantage within the infected host during chronic infection. The negative HBeAg mutants have a biological advantage over the wild strain of the virus, so these strains are selected throughout evolution.

In the general population, the wild type of HBV still predominates over the mutant forms.
Also, the mutated form of the stop codon in the “precore” region is associated with fulminant hepatitis and is much more frequently found in HBV genotype D.

In such cases, the therapy of choice is lamivudine, which acts on the dominant variant mutated into “precore”.

It is known that IFN has a potent antiviral action only on the wild virus strain. Therefore, the therapeutic decision must be made individually, only after a molecular virology diagnosis, in order to have a sustained virological response, in fact, a therapeutic success.

Specializations in immunology

  • July 1997 – London UK, Chelsea & Westminster Hospital, Dept. of Immunology, Prof. Frances M. Goth;
  • July – 1997 – London, UK, Hammersmith Hospital RPMS, Imperial College of Science Technology and Medicine – Dept. of Immunology and Virology – Infectious Diseases Dept.: Dr. Kate N. Ward;
  • July 1997 – London, UK, Public Health Laboratory Service PHLS – Virus Reference Laboratory – Dr. CG Teo – Specialization in Molecular Biology, Virology – Molecular Diagnosis – acquisition of the gene amplification technique for HCV and HBV viruses. Acquiring genotyping techniques.
  • July 1998 – London, UK, Hammersmith Hospital RPMS, Imperial College of Science Technology and Medicine – Dept. of Immunology and Virology – Infectious Diseases Dept.: Dr. Kate N. Ward;

Long-term international scientific collaborations

  1. December 1999  – Anti-viral therapy? in chronic hepatitis B and C: patient selection, therapeutic regimens, clinical, paraclinical and histopathological parameters that are followed in the evolution of the viral infection – collaboration carried out in January at the Institute for the Study of the Liver, Kaplan, Rehovot, Israel Dr. Yoav Lurie.
  2. February 2000  – Incidence of hepatocellular carcinoma after interferon therapy in patients with chronic hepatitis B and C. Sequencing of HCV and HBV, with the study of HBV and HCV mutations in Romanian patients. Cloning studies of these viruses. Molecular biology techniques for the detection of HCV RNA and HBV DNA in serum and liver tissue. Research and clinical internship – Hepatology – Virology between 20.01-31.03.2000, carried out at the Royal Free Hospital, University College of London, with Dr. Tim Harrison and Prof. Geofrey Dusheiko. ‘Core Promoter Mutations in Patients with Chronic hepatitis B and Hepatocellular Carcinoma in Bucharest, Romania research project with the collaboration of Dr. Tim Harrison.
  3. July 2002  – Collaboration with The Anthony Nolan Bone Marrow Trust, Royal Free Hospital, Hampstead, London, UK, with Joyce Beo PhD, John O’Shea in the field of Bone Marrow Transplantation.
  4. June 2003  – Collaboration with the Immunogenetics Center of the National Medical Center, from Budapest, Hungary – Prof. Gyozo Petranyi – within the bilateral marrow transplant program. Creation of the national register of marrow donors in the Fundeni Clinical Institute.
  5. May 2004  – Collaboration with the Immunology and Histocompatibility Center “Evangelismos” Hospital, Athens, Greece – Dr. Chryssa Papasteriades, EFI responsible for the 8th European region, of which Romania is also a part.
  6. October 2004  – collaboration with Prof. Ilias Doxiadis from the Eurotransplant Reference Laboratory in Leiden, Holland in the field of Histocompatibility; dialogue regarding the accreditation process of Romanian Histocompatibility laboratories.
  7. October 2005  – Collaboration with “Central Laboratory of Clinical Immunology – University Hospital Alexandrovska” from Bulgaria – Prof. Dr. Elisaveta Naumova, Dr. Milena Ivanova, in the field of HLA polymorphism and cytokine genes in the elderly, results presented at the 14th meeting IHIW of the European Federation of Immunogenetics during November 29 – December 3, 2005, Melbourne, Australia.
  8. November 2005  – Collaboration with UKNEQAS (United Kindom National External Quality Assessment Schemes) – professional association of external quality control, from Great Britain, in the field of medical education and quality control in the field of Molecular Biology, Immunogenetics, Transplant Immunology, Tumor Immunology and Virology.

The laboratory was accredited by the European Federation of Immunogenetics (EFI) in 2006.