Bulgurcu, Alihan
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BULGURCU, ALIHAN
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alihan.bulgurcu@ieu.edu.tr
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15.06. Medical Laboratory Techniques
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3GOOD HEALTH AND WELL-BEING
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4QUALITY EDUCATION
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57
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9
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13
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1.50
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2.17
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| Journal | Count |
|---|---|
| Mikrobiyoloji Bulteni | 2 |
| BMC Microbiology | 1 |
| FEBS Open Bio | 1 |
| Mıkrobıyolojı Bultenı | 1 |
| Turkısh Journal of Bıochemıstry-Turk Bıyokımya Dergısı | 1 |
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Now showing 1 - 6 of 6
Article Citation - WoS: 2Citation - Scopus: 2Analysis of Nucleotide Changes in Rt-Pcr Primer/Probe Binding Regions in Sars-Cov Isolates Reported From Turkey(Ankara Microbiology Soc, 2021-07-16) Demir, Ayse Banu; Bulgurcu, Alihan; Appak, Ozgur; Sayiner, Ayca ArzuThe SARS-CoV-2 virus, which caused the COVID-19 epidemic, caused more than 55 million cases and nearly 1.5 million deaths worldwide. For the microbiological diagnosis of the disease, the most valid method is detecting the presence of the viral genome by real-time reverse transcription polymerase chain reaction (rRT-PCR). However, due to the nature of the RNA viruses, frequent mutations may affect the sensitivity of the analyses made on the genetic material of the virus, such as PCR. In this study, we aimed to investigate the mutations in the primer-probe binding regions of the rRT-PCR panels used in COVID-19 diagnosis. SARS-CoV-2 whole genome sequence data (n= 194) isolated from COVID-19 cases in Turkey and uploaded on GISAID database from the centers in Istanbul (n= 78), Ankara (n= 58), Kars (n= 47), Bursa (n= 2), Adiyaman (n= 2), Erciyes (n= 1) and Kocaeli (n= 1) between March 17-September 14, 2020 were analyzed. In order to determine the nucleotide changes, SARS-CoV-2 sequences from Turkey were compared to the reference genome sequence (NC_045512.1) present in GenBank website. The constructed data set was aligned using the MAFFT program and was checked manually if the sequences were in the same frame by using the AliView program. Primer-probe binding sites of the thirteen SARS-CoV-2 rRT-PCR panels from seven different institutes (US CDC, China CDC, Charite CDC, Pasteur, HKU, Thailand, NIID) that are being used in COVID-19 diagnosis were evaluated in terms of nucleotide changes within the corresponding regions compared to the reference genome. Sequence diversities in the viral genomes were determined via positional nucleotide numerical calculator and entropy calculator modules and nucleotide and entropy changes in primer-probe binding regions for each rRT-PCR panel were examined. Among thirteen different primer-probe panels, nucleotide changes in the target regions of the seven primer-probe panels were determined. When viral sequences with nucleotide changes in the primer-probe binding regions were examined, the most common changes were observed in the China CDC N-forward primer and US CDC N3-forward primer binding regions. It is important that the kits to be used as diagnostic tests are designed specific to the regions with less nucleotide changes. Nucleotide changes may not be critical for DNA amplification for most PCR panels, but should be carefully monitored as they may affect the sensitivity of the assay. If the risk of alteration of the designed region is high, the primer - probe binding sites should be checked frequently and updated when necessary.Conference Object Development of Reference Materials for the Validation of Influenza A/B and RSV Nucleic Acid Detection Kits Using Digital PCR(Wiley, 2025) Bulgurcu, Alihan; Sayiner, A. A.Article İnfluenza A/B Virüs ve RSV Validasyon Standartlarının Dijital PCR ile Kantitasyonu(Ankara Microbiology Soc, 2025-07-26) Sayıner, Ayca Arzu; Bulgurcu, AlıhanMikrobiyolojik tanı laboratuvarlarında kullanılacak tanı testleri için kantitatif standartların kullanıldığı yöntem doğrulama (verification) veya geçerli kılma (validation) çalışmaları gereklidir. Nükleik asit testlerinde sentetik nükleik asit veya plazmit yerine tam virüs içeren standartların kullanılması; ekstraksiyon, revers transkripsiyon ve amplifikasyonu içerecek şekilde tanı testinin tüm basamaklarının gerçek yaşam koşullarında değerlendirilmesini sağlar. Solunum yolu virüsleri için nükleik asit testlerine yönelik ticari kantitatif standart materyaller sınırlıdır. Bu çalışmada; influenza A virüs (infA), influenza B virüs (infB) ve respiratuvar sinsityal virüs (RSV) için dijital polimeraz zincir reaksiyonu [digital polymerase chain reaction (dPCR)] kullanılarak, kantitatif nükleik asit standartları geliştirilmesi amaçlanmıştır. Çalışmada; RSV, infA, infB RNA pozitif olduğu bilinen nazofarengeal sürüntü örneklerinin havuzlanmasıyla hazırlanan örneklerdeki viral nükleik asit miktarı, ticari primer/prob setleri (Qiagen, Almanya) kullanılarak dPCR (QIAcuity, Qiagen) yöntemiyle belirlenmiştir. Nükleik asit ekstraksiyonu, ticari bir kit (Xi’an Tianlong Science&Technology Co, Çin) kullanılarak yapılmıştır. dPCR yönteminin infA, infB ve RSV için analitik duyarlılık (LoD) ve kantitasyon alt sınırı (LoQ), çalışma içi ve çalışmalar arası tekrarlanabilirliği ve doğrusallığı belirlenmiştir. dPCR ile çalışılan örnekler, kantitatif revers transkripsiyon gerçek zamanlı [quantitative reverse transcription realtime (qrRT)] PCR (qRT-PCR) ile de çalışılarak Ct değerleri belirlenmiştir. Ct değerleri ile dPCR-kantitasyon sonuçları arasındaki ilişki lineer regresyon ile değerlendirilmiştir. İstatistiksel analiz GraphPad Prism 10.4.0 (GraphPad, ABD) ve Excel Analysis ToolPak kullanılarak yapılmıştır. İnfA, infB ve RSV için dPCR yönteminin LoD değerleri sırasıyla 93.75, 15.59 ve 26.23 kopya/mL olarak belirlenmiştir. dPCR yönteminin çalışma içi tekrarlanabilirliği (varyasyon katsayısı, %CV), düşük viral yükü olan örneklerde daha yüksek olmak üzere 0.06-7.97 arası saptanmıştır. Çalışmalar arası tekrarlanabilirlik 0.73-5.41 olarak bulunmuştur. İnfA ve infB için 3-4 log10, RSV için 7 log10 aralığında dilüsyonlar ile yapılan doğrusallık analizinde her üç virüs için de r 2≥ 0.99 olarak bulunmuştur. dPCR ile ölçülen konsantrasyonların, qRT-PCR Ct sonuçları ile korele olduğu saptanmıştır. dPCR ile qRT-PCR testlerinin çalışma içi ve çalışmalar arası tekrarlanabilirlik sonuçları karşılaştırıldığında, dPCR’nin %CV değerinin anlamlı olarak daha düşük olduğu saptanmıştır (p= 0.0312). Çalışma sonuçları dPCR yönteminin, kantitatif nükleik asit standartları elde etmede tekrarlanabilirliği yüksek ve güvenilir bir yöntem olduğunu göstermiştir. Elde edilen kantitatif standartlar ile viral yük belirlemeye yönelik tanı yöntemleri geliştirmek ve/veya bu tür testlerin yöntem onayı analizlerini yapmak mümkündür. Sonuç olarak çalışmada, havuzlanmış hasta örnekleri kullanılarak dPCR yöntemiyle infA, infB ve RSV için güvenilir kantitatif nükleik asit standartlar elde edilmiş ve dPCR yönteminin performans analizleri gerçekleştirilmiştir. Bu çalışma, dPCR ile kantitatif viral nükleik asit standartlarının üretimine bir örnek olmuştur.Article Validation of a Multiplex Qrt-PCR Assay for the Detection of RSV, Influenza A/B Virus and SARS-CoV(BMC, 2026-01-03) Bulgurcu, Alihan; Sayiner, Ayca ArzuThe significant burden of viral respiratory diseases necessitates rapid detection of key pathogens. Simultaneous testing for SARS-CoV-2, RSV, and influenza A/B as an initial step, followed by broader panels as needed, offers a cost-effective diagnostic strategy. This study aimed to validate a new commercial multiplex qRT-PCR assay (Diagnovital (R) RTA Laboratories, Turkey) for the simultaneous detection of these viruses. Analytical sensitivity was determined using Probit regression analysis on serial dilutions (10(5)-10(1) copies/ml) for each target virus. Specificity was evaluated with 120 negative samples and 32 positive samples for non-target respiratory viruses. External quality control panels and clinical specimens positive for RSV (n = 39), influenza A/B (n = 71), SARS-CoV-2 (n = 64) were used for accuracy testing. Intra- and inter-assay precision were analyzed using samples near the limit of detection. The performance was compared to routine diagnostic tests. The assay's analytical sensitivity was 420.7, 296.7, 368.6, 1362.6, and 1459.7 copies/ml for SARS-CoV-2 Alpha and Omicron variants, influenza A, influenza B, and RSV, respectively. Analytical specificity was 100%, and precision showed CV% < 5. Detection rates for SARS-CoV-2, influenza A, influenza B, and RSV were 100%, 95.1%, 97.5%, and 94.8%, respectively, with false negatives occurring in samples with Ct > 33. Comparative analysis showed high correlations between assays, with strong agreement (Cohen's kappa ranging from 0.861 to 1). These findings demonstrate the clinical applicability of the Diagnovital (R) assay, though false negatives may occur in low-concentration samples.Article Citation - WoS: 7Citation - Scopus: 11The Effect of Virtual Laboratory Simulations on Medical Laboratory Techniques Students' Knowledge and Vocational Laboratory Education(Walter De Gruyter Gmbh, 2022-08-01) Keles, Didem; Bulgurcu, Alihan; Demir, Esra Feyzioglu; Şemin, Makbule İlgi; Feyzioğlu Demir, Esra; Şemin, Ilgi Makbule; Feyzioğlu-demir, EsraObjectives Virtual laboratory simulations (VLSs) are computer-based tools that offer unlimited application options in scientific, medical, and engineering fields. The aim of this study was to evaluate whether VLSs are efficient learning tools and how these simulations can be integrated into laboratory practice in medical laboratory education. Methods In this pre-test/post-test control group study, 32 volunteers were randomly assigned to either experimental or control groups. The experimental group performed laboratory simulations based on biochemistry and microbiology and then completed a self-report survey to evaluate their satisfaction and beliefs about simulations. Results In the experimental group, post-test scores of each simulation were significantly elevated compared to pre-test scores; however, pre- and post-test scores of control group were statistically the same. The experimental group agreed that these simulations should be applied before theoretical lectures and laboratory practices. They also highlighted that translating from English to their native language creates difficulties in applying and understanding the simulation. Conclusions We emphasized that VLSs are excellent learning tools that increase not only the knowledge but also the self-motivation and focus of the students. Based on feedbacks, native language options are necessary to enable the students to achieve equality of opportunity in education.Article Analysis of Nucleotide Changes in Rt-Pcr Primer/Probe Binding Regions in Sars-Cov-2 Isolates Reported from Turkey(Ankara Microbiology Society, 2021-07-16) Sayıner, Ayça Arzu; Appak, Özgür; Demir, Ayse Banu; Bulgurcu, AlihanThe SARS-CoV-2 virus, which caused the COVID-19 epidemic, caused more than 55 million cases and nearly 1.5 million deaths worldwide. For the microbiological diagnosis of the disease, the most valid method is detecting the presence of the viral genome by real-time reverse transcription polymerase chain reaction (rRT-PCR). However, due to the nature of the RNA viruses, frequent mutations may affect the sensitivity of the analyses made on the genetic material of the virus, such as PCR. In this study, we aimed to investigate the mutations in the primer-probe binding regions of the rRT-PCR panels used in COVID-19 diagnosis. SARS-CoV-2 whole genome sequence data (n= 194) isolated from COVID-19 cases in Turkey and uploaded on GISAID database from the centers in Istanbul (n= 78), Ankara (n= 58), Kars (n= 47), Bursa (n= 2), Adiyaman (n= 2), Erciyes (n= 1) and Kocaeli (n= 1) between March 17-September 14, 2020 were analyzed. In order to determine the nucleotide changes, SARS-CoV-2 sequences from Turkey were compared to the reference genome sequence (NC_045512.1) present in GenBank website. The constructed data set was aligned using the MAFFT program and was checked manually if the sequences were in the same frame by using the AliView program. Primer-probe binding sites of the thirteen SARS-CoV-2 rRT-PCR panels from seven different institutes (US CDC, China CDC, Charite CDC, Pasteur, HKU, Thailand, NIID) that are being used in COVID-19 diagnosis were evaluated in terms of nucleotide changes within the corresponding regions compared to the reference genome. Sequence diversities in the viral genomes were determined via positional nucleotide numerical calculator and entropy calculator modules and nucleotide and entropy changes in primer-probe binding regions for each rRT-PCR panel were examined. Among thirteen different primer-probe panels, nucleotide changes in the target regions of the seven primer-probe panels were determined. When viral sequences with nucleotide changes in the primer-probe binding regions were examined, the most common changes were observed in the China CDC N-forward primer and US CDC N3-forward primer binding regions. It is important that the kits to be used as diagnostic tests are designed specific to the regions with less nucleotide changes. Nucleotide changes may not be critical for DNA amplification for most PCR panels, but should be carefully monitored as they may affect the sensitivity of the assay. If the risk of alteration of the designed region is high, the primer - probe binding sites should be checked frequently and updated when necessary.
