Genotyping clinical cytomegalovirus isolates in solid-organs-transplant recipients

Cover Page


Cite item

Full Text

Abstract

Cytomegalovirus infection remains one of the leading problems in contemporary healthcare. It belongs to socially and economically significant infections with a high incidence both in children and adults, characterized by polymorphic clinical manifestations and a variety of routes and factors for infection transmission. CMV infection of blood and organ recipients is a serious problem. It should be noted that, despite the great medical and social significance, in the Russian Federation there is no system of CMV epidemiological surveillance and control as it was traditionally developed for other topical infections. The aim of this study is to search for optimal method of cytomegalovirus (CMV) genotyping and estimate genotypic diversity of CMV isolates in Russia for patients underwent solid organ transplantation. The research presents the data after examining blood samples, leukocytes, saliva, urine and lacrimal discharge collected from 160 patients at the Transplantation Department of the Privolzhsky District Medical Center of the FMBA, aged 22 to 64 years, after liver and kidney transplantation. Molecular biological and serological methods were used for testing. Genotyping was carried out by the NGS sequencing of CMV DNA fragments. A high prevalence of CMV was found in patients undergoing solid organ transplantation. For 41.8±3.8% patients, cytomegalovirus DNA was detected in saliva and urine samples, and for 18.1±3.04% of them — in blood samples. In 98.8±3.2% of patients, the diagnosis of Cytomegalovirus infection (CMVI) was confirmed serologically. Based on a summary of reported data, estimation of various methodological approaches for genotyping of clinical CMV isolates was carried out. As a result, a typing option based on genotype determining for two variable genes UL55 (gB) and UL73 (gN) was selected. The spectra and proportional distribution of gB and gN CMV genotypes circulating among adults were determined. It was found that genotype prevalence in the group of patients who underwent solid organ transplantation was as follows: gB2, gN4c, gN4a, gN1. In some cases, a mixed infection was found due to the association of two and three CMV genotypes. The performed phylogenetic analysis of UL55 and UL73 gene nucleotide sequences indicates the genetic heterogeneity found for Russia-wide CMV isolates from in adult patients in the risk group.

About the authors

O. E. Vankova

Blokhina Scientific Research Institute of Epidemiology and Microbiology

Author for correspondence.
Email: voe0@mail.ru
ORCID iD: 0000-0002-9838-1133

Olga E. Vankova - Senior Researcher, Laboratory of Metagenomics and Molecular Indication of Pathogens, Blokhina Scientific Research Institute of Epidemiology and Microbiology.

603950, Nizhny Novgorod, Malaya Yamskaya str., 71.

Phone: +7 920 022-63-80.

Russian Federation

N. F. Brusnigina

Blokhina Scientific Research Institute of Epidemiology and Microbiology

Email: nfbrusnigina@yandex.ru
ORCID iD: 0000-0003-4582-5623

PhD (Medicine), Head of the Laboratory of Metagenomics and Molecular Indication of Pathogens, Blokhina Scientific Research Institute of Epidemiology and Microbiology.

603950, Nizhny Novgorod, Malaya Yamskaya str., 71.

Russian Federation

References

  1. Barbi M., Binda S., Caroppo S., Calvario A., Germinario C., Bozzi A., Tanzi M.L., Veronesi L., Mura I., Piana A., Solinas G., Pugni L., Evilaqua G., Mosca F. CMV gB genotypes and outcome of vertical transmission: study on dried blood spots of congenitally infected babies. J. Clin. Virol., 2001, vol. 21, no. 1, pp. 75–79. doi: 10.1016/s1386-6532(00)00188-8
  2. Cannon M.J., Schmid D.S., Hyde T.B. Review of cytomegalovirus seroprevalence and demographic characteristics associated with infection. Rev. Med. Virol., 2010, vol. 20, no. 4, pp. 202–213. doi: 10.1002/rmv.655
  3. Chen H.P., Lin J.C., Yang S.P., Lan Y.C., Weng W.S., Tsai C.H., Ho D.M., Liu C.Y., Cho W.L., Chan Y.J. The type-2 variant of human cytomegalovirus glycoprotein N (gN-2) is not the rarest in the Chinese population of Taiwan: influence of primer design. J. Virol. Methods, 2008, vol. 151, pp. 161–164. doi: 10.1016/j.jviromet.2008.03.018
  4. Chou S.W., Dennison K.M. Analysis of interstrain variation in cytomegalovirus glycoprotein B sequences encoding neutralization related epitopes. J. Infect. Dis., 1991, vol. 163, no. 6, pp. 1229–1234. doi: 10.1093/infdis/163.6.1229
  5. Ciotti M., Cella E., Rittà M., Ciccozzi M., Cavallo R., Perno C.F., Costa C. Cytomegalovirus glycoprotein B genotype distribution in Italian transplant patients. Intervirology, 2017, vol. 60, no. 4, pp. 165–170. doi: 10.1159/000486593
  6. De Albuquerque D.M., Costa S.C. Genotyping of human cytomegalovirus using non-radioactive single-strand conformation polymorphism (SSCP) analysis. J. Virol. Methods., 2003, vol. 9, no. 110 (1), pp. 25–28. doi: 10.1016/s0166-0934(03)00094-6
  7. De Vries J.J., Wessels E., Korver A.M., van der Eijk A.A., Rusman L.G., Kroes A.C., Vossen A.C. Rapid genotyping of cytomegalovirus in dried blood spots by multiplex real-time PCR assays targeting the envelope glycoprotein gB and gH genes. J. Clin. Microbiol., 2012, vol. 50, no. 2, pp. 232–237. doi: 10.1128/JCM.05253-11
  8. Dieamant D.C., Bonon S.H., Peres R.M., Costa C.R., Albuquerque D.M., Miranda E.C., Aranha F.J., Oliveira-Duarte G., Fernandes V.C., De Souza C.A., Costa S.C., Vigorito A.C. Cytomegalovirus (CMV) genotype in allogeneic hematopoietic stem cell transplantation. BMC Infect. Dis., 2013, vol. 10, no. 13: 310. doi: 10.1186/1471-2334-13-310
  9. Görzer I., Guelly С., Trajanoski S., Puchhammer-Stöckl E. Deep sequencing reveals highly complex dynamics of human cytomegalovirus genotypes in transplant patients over time. J. Virol., 2010, vol. 84, no. 14, pp. 7195–7203. doi: 10.1128/JVI.00475-10
  10. Grosjean J., Hantz S., Cotin S., Baclet M.C., Mengelle C., Trapes L., Virey B., Undreiner F., Brosset P., Pasquier C., Denis F., Alain S. Direct genotyping of cytomegalovirus envelope glycoproteins from toddler’s saliva samples. J. Clin. Virol., 2009, vol. 46, no. 4, pp. 43–48. doi: 10.1016/j.jcv.2009.08.018
  11. Khalafkhany D., Makhdoomi K., Taghizadeh Afshari A., Motazakker M. Prevalence and genotype distribution of cytomegalovirus UL55 sequence in renal transplant recipients in north west of Iran. J. Med. Virol., 2016, vol. 88, no. 9, pp. 1622–1627. doi: 10.1002/jmv.24509
  12. Larkin M.A., Blackshields G., Brown N.P., Chenna R., McGettigan P.A., McWilliam H., Valentin F., Wallace I.M., Wilm A., Lopez R., Thompson J.D., Gibson T.J., Higgins D.G. Clustal W and Clustal X version 2.0. Bioiformatics, 2007, vol. 23, no. 21, pp. 2947–2948. doi: 10.1093/bioinformatics/btm404
  13. Lisboa L.F., Tong Y., Kumar D., Pang X.L., Asberg A., Hartmann A., Rollag H., Jardine A.G., Pescovitz M.D., Humar A. Analysis and clinical correlation of genetic variation in cytomegalovirus. Transpl. Infect. Dis., 2012, vol. 14, no. 2, pp. 132–140. doi: 10.1111/j.1399-3062.2011.00685.x
  14. Manuel O., Asberg A., Pang X., Rollag H., Emery V.C., Preiksaitis J.K., Kumar D., Pescovitz M.D., Bignamini A.A., Hartmann A., Jardine A.G., Humar A. Impact of genetic polymorphisms in cytomegalovirus glycoprotein B on outcomes in solid-organ transplant recipients with cytomegalovirus disease. Clin. Infect. Dis., 2009, vol. 15, no. 49 (8), pp. 1160–1166. doi: 10.1086/605633
  15. Murthy S., Hayward S., Wheelan S., Forman M.S., Ahn J.H., Pass R.F., Arav-Boger R. Detection of a single identical cytomegalovirus (CMV) strain in recently seroconverted young women. PLoS One, 2011, vol. 10, no. 6 (1), pp. 149–159. doi: 10.1371/journal.pone.001594
  16. Navarro-Rodríguez V., Herrera-Munoz A., Castro A., Ramos-Esquivel A. Risk factors for cytomegalovirus disease in seropositive renal transplant recipients; a single-center case-controlled study. J. Nephropathol., 2017, vol. 6, no. 3, pp. 240–247. doi: 10.15171/jnp.2017.39
  17. Pignatelli S. Recent knowledge on the linkage of strain specific genotypes with clinical manifestations of human citomegalovirus disease. Recenti. Prog. Med., 2011, vol. 102, no. 1, pp. 5–10.
  18. Pignatelli S., Dal Monte Р. Epidemiology of human cytomegalovirus strains through comparison of methodological approaches to explore gN variants. New Microbiol., 2009, vol. 32, no. 1, pp. 1–10.
  19. Roubalova K. Genetic variability of cytomegalovirus glycoprotein O in hematopoietic stem cell transplant recipients. Transpl. Infect. Dis., 2011, vol. 13, no. 3, pp. 237–243. doi: 10.1111/j.1399-3062.2011.00625.x
  20. Roubalová K.О., Strunecký S., Zufanová S., Procházka B., Vitek A. Genotyping of viral glycoprotein B (gB) in hematopoietic stem cell transplant recipients with active cytomegalovirus infection: analysis of the impact of gB genotypes on the patients’ out-come. Epidemiol. Mikrobiol. Imunol., 2010, vol. 59, no. 2, pp. 92–99.
  21. Shepp D.H., Match M.E., Lipson S.M., Pergolizzi R.G. A fifth human cytomegalovirus glycoprotein B genotype. Res. Virol., 1998, vol. 149, no. 2, pp. 109–114. doi: 10.1016/s0923-2516(98)80086-1
  22. Torii Y., Yoshida S., Yanase Y., Mitsui T., Horiba K., Okumura T., Takeuchi S., Suzuki T., Kawada J.I., Kotani T., Yamashita M., Ito Y. Serological screening of immunoglobulin M and immunoglobulin G during pregnancy for predicting congenital cytomegalovirus infection. BMC Pregnancy Childbirth, 2019, vol. 19, no. 1: 205. doi: 10.1186/s12884-019-2360-1
  23. Varghese J., Subramanian S., Reddy M.S., Shanmugam N., Balajee G., Srinivasan V., Venkataraman J., Mohamed R. Seroprevalence of cytomegalovirus in donors and opportunistic viral infections in liver transplant recipients. Indian J. Med. Res., 2017, vol. 145, no. 4, pp. 558–562.
  24. Wu X.J., Wang Y., Zhu Z., Xu Y., He G., Han Y., Tang X., Fu Z., Qiu H., Sun A., Wu D. The correlation of cytomegalovirus gB genotype with viral DNA load and treatment time in patients with CMV infection after hematopoietic stem cell transplantation. Zhonghua Xue Ye Xue Za Zhi, 2013, vol. 34, no. 2, pp. 109–112.
  25. Xia C.S., Zhang Z. Analysis of human cytomegalovirus glycoprotein N genotypes in Chinese hematopoietic stem cell transplant recipients. Arch. Virol., 2011, vol. 156, no. 1, pp. 17–23. doi: 10.1007/s00705-010-0811-0

Supplementary files

Supplementary Files
Action
1. JATS XML
Download

Copyright (c) 2022 Vankova O.E., Brusnigina N.F.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № ФС 77 - 64788 от 02.02.2016.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies