Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples

Aim The two most common human polyomaviruses are the BK (BKV) and JC viruses (JCV). Diseases associated with polyomavirus usually occur in cases of severe cellular immunosuppression. BKV and JCV can cause many diseases, especially if they are reactivated in an immunosuppressed host. The aim of this...

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Main Authors: Mehmet Özdemir, Uğur Ayan, Murat Şevik
Format: Article
Language:English
Published: Thieme Medical and Scientific Publishers Pvt. Ltd. 2020-08-01
Series:Journal of Laboratory Physicians
Subjects:
Online Access:http://www.thieme-connect.de/DOI/DOI?10.1055/s-0040-1716603
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spelling doaj-4b087a28df2144009f6aaad8dd8af3232020-11-25T03:32:23ZengThieme Medical and Scientific Publishers Pvt. Ltd.Journal of Laboratory Physicians0974-27270974-78262020-08-01120207908310.1055/s-0040-1716603Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical SamplesMehmet Özdemir0Uğur Ayan1Murat Şevik2Department of Medical Microbiology, Division of Medical Virology, Meram Medical Faculty, Necmettin Erbakan University, Konya, TurkeyMedical Microbiology Laboratory, Istanbul Medeniyet University, Göztepe Training and Research Hospital, Istanbul, TurkeyDepartment of Virology, Veterinary Faculty, Mustafa Kemal University, Antakya, TurkeyAim The two most common human polyomaviruses are the BK (BKV) and JC viruses (JCV). Diseases associated with polyomavirus usually occur in cases of severe cellular immunosuppression. BKV and JCV can cause many diseases, especially if they are reactivated in an immunosuppressed host. The aim of this study is to compare and evaluate the results of real-time polymerase chain reaction (PCR) methods targeting the small and large T gene regions of the viral genome, considering polymorphisms occurring in the viral genome of BKV and JCV. Materials and Methods Urinary specimens of 82 patients were taken from immunosuppressed patient and sent to molecular microbiology laboratory of Meram Medical Faculty. The small t gene was investigated using a commercial kit (LightMix, Roche) by real-time PCR method. Large T gene was investigated by using the optimized in-house real-time PCR method. Sequence analysis was accepted as the standard method. Results BKV positivity was detected in 9 samples and JCV positivity in 61 samples by real-time PCR method specific to small t gene region; BKV positivity in 21 samples and JCV positivity in 67 samples were determined by real-time PCR method specific to the large T gene region. Statistically, there was a significant difference for BKV, but not significant difference for JCV detection between the two methods. Conclusion Different polymorphisms in the target gene regions were responsible for the different outcomes obtained from this study. With this sensitivity and specificity, in-house PCR method which we used is a candidate for routine diagnosis.http://www.thieme-connect.de/DOI/DOI?10.1055/s-0040-1716603bk virusjc virusreal-time pcr
collection DOAJ
language English
format Article
sources DOAJ
author Mehmet Özdemir
Uğur Ayan
Murat Şevik
spellingShingle Mehmet Özdemir
Uğur Ayan
Murat Şevik
Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
Journal of Laboratory Physicians
bk virus
jc virus
real-time pcr
author_facet Mehmet Özdemir
Uğur Ayan
Murat Şevik
author_sort Mehmet Özdemir
title Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
title_short Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
title_full Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
title_fullStr Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
title_full_unstemmed Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples
title_sort comparative evaluation of in-house and commercial real-time pcr methods for the detection of the bk and jc viruses in clinical samples
publisher Thieme Medical and Scientific Publishers Pvt. Ltd.
series Journal of Laboratory Physicians
issn 0974-2727
0974-7826
publishDate 2020-08-01
description Aim The two most common human polyomaviruses are the BK (BKV) and JC viruses (JCV). Diseases associated with polyomavirus usually occur in cases of severe cellular immunosuppression. BKV and JCV can cause many diseases, especially if they are reactivated in an immunosuppressed host. The aim of this study is to compare and evaluate the results of real-time polymerase chain reaction (PCR) methods targeting the small and large T gene regions of the viral genome, considering polymorphisms occurring in the viral genome of BKV and JCV. Materials and Methods Urinary specimens of 82 patients were taken from immunosuppressed patient and sent to molecular microbiology laboratory of Meram Medical Faculty. The small t gene was investigated using a commercial kit (LightMix, Roche) by real-time PCR method. Large T gene was investigated by using the optimized in-house real-time PCR method. Sequence analysis was accepted as the standard method. Results BKV positivity was detected in 9 samples and JCV positivity in 61 samples by real-time PCR method specific to small t gene region; BKV positivity in 21 samples and JCV positivity in 67 samples were determined by real-time PCR method specific to the large T gene region. Statistically, there was a significant difference for BKV, but not significant difference for JCV detection between the two methods. Conclusion Different polymorphisms in the target gene regions were responsible for the different outcomes obtained from this study. With this sensitivity and specificity, in-house PCR method which we used is a candidate for routine diagnosis.
topic bk virus
jc virus
real-time pcr
url http://www.thieme-connect.de/DOI/DOI?10.1055/s-0040-1716603
work_keys_str_mv AT mehmetozdemir comparativeevaluationofinhouseandcommercialrealtimepcrmethodsforthedetectionofthebkandjcvirusesinclinicalsamples
AT ugurayan comparativeevaluationofinhouseandcommercialrealtimepcrmethodsforthedetectionofthebkandjcvirusesinclinicalsamples
AT muratsevik comparativeevaluationofinhouseandcommercialrealtimepcrmethodsforthedetectionofthebkandjcvirusesinclinicalsamples
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