Determination of Genotypes in Cytomegalovirus (CMV) Strains Obtained from Pediatric and Adult Immunocompromised Patients

Abstract

Background and Purpose: Cytomegalovirus causes asymptomatic disease in individuals with normal immune system; and leads to serious complications in immunocompromised individuals and fetus. In CMV, gB is the most studied glycoprotein in terms of genotyping. Up to now, four different gB genotypes (gB 1-4) of CMV have been identified. In this study, it was aimed to determine the genotypes of CMV strains obtained from patients with immune deficiency. Methods: Twenty children and 29 adults, 49 patients who were followed in the Department of Adult Hematology and Pediatric Hematology were included in the study. DNA isolation was performed from samples with CMV DNA level of 1000 IU / ml and above, and 474 bp region from the gB region of the virus was amplified by nested PCR. This region was sequenced by the Sanger (ABI 3500 Prism) sequencing. Next generation sequencing (NGS) method was applied to the samples that CMV genotype could not be determined by Sanger sequencing. Results: Distribution of CMV genotypes of patients determined by Sanger sequencing; while it was determined as 18/49 (36.7%) type 1, 5/49 (10.2%) type 2, 5/49 (10.2%) type 3 and 1/49 (2%) type 4; 14/49 (28.5%) of them were found as mixed genotypes. CMV genotype could not be determined in 6 patients by Sanger sequencing and CMV genotype of these 6 patients were found as mixed genoype by NGS. Mixed genotype was detected in 20 (40.9%) of 49 patients, in total by Sanger sequencing and NGS. Conclusion: It is remarkable that the most common genotype is mixed genotype. It was concluded that it is appropriate to study the next generation sequencing method in patients whose CMV genotype cannot be determined by Sanger sequencing.

Keywords

• Cytomegalovirus
• genotype
• sequencing
• NGS

References

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