Bu çalışma, Ulusal Sağlık Enstitüleri Klinik Merkezi'nin Intramural Araştırma Programı tarafından desteklenmiştir. İçerik yalnızca yazarların sorumluluğundadır ve Ulusal Sağlık Enstitüleri'nin resmi görüşlerini temsil etmemektedir.

Temiz Oda Ortamları için Mikrobiyal Kontrol ve İzleme Stratejileri

<ol>
	<li>Cundell, T., Atkins, J. W., Lau, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sterility+testing+for+hematopoietic+stem+cells.">Sterility testing for hematopoietic stem cells.</a> Journal of Clinical Microbiology. , (2023).</li><li>United States Food and Drug Administration. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidance+for+Industry.+Sterile+Drug+Products+Produced+by+Aseptic+Processing+-+Current+Good+Manufacturing+Practice.">Guidance for Industry. Sterile Drug Products Produced by Aseptic Processing - Current Good Manufacturing Practice.</a> United States Food and Drug Administration. , (2004).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=USP&lt;71&gt;+Sterility+Tests+in+USP43-NF38.">USP&lt;71&gt; Sterility Tests in USP43-NF38.</a> United States Pharmacopeia - National Formulary. , (2022).</li><li>England, M. R., Stock, F., Gebo, J. E. T., Frank, K. M., Lau, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+evaluation+of+compendial+USP&lt;71&gt;,+BacT/Alert+Dual-T,+and+Bactec+FX+for+detection+of+product+sterility+testing+contaminants.">Comprehensive evaluation of compendial USP&lt;71&gt;, BacT/Alert Dual-T, and Bactec FX for detection of product sterility testing contaminants.</a> Journal of Clinical Microbiology. 57 (2), 01548 (2019).</li><li>Gebo, J. E. T., East, A. D., Lau, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+side-by-side+comparison+of+clinical+versus+current+good+manufacturing+practices+(cGMP)+microbiology+laboratory+requirements+for+sterility+testing+of+cellular+and+gene+therapy+products.">A side-by-side comparison of clinical versus current good manufacturing practices (cGMP) microbiology laboratory requirements for sterility testing of cellular and gene therapy products.</a> Clinical Microbiology Newsletter. 43 (21), 181-191 (2021).</li><li>Gebo, J. E. T., Lau, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sterility+testing+for+cellular+therapies:+What+is+the+role+of+the+clinical+microbiology+laboratory.">Sterility testing for cellular therapies: What is the role of the clinical microbiology laboratory.</a> Journal of Clinical Microbiology. 58 (7), 01492 (2020).</li><li>International Organization for Standardization. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ISO+14644-1:2015+Cleanrooms+and+associated+controlled+environments+-+Part+1:+Classification+of+air+cleanliness+by+particle+concentration.">ISO 14644-1:2015 Cleanrooms and associated controlled environments - Part 1: Classification of air cleanliness by particle concentration.</a> International Organization for Standardization. , (2015).</li><li>Parenteral Drug Association. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PDA+Technical+Report+No.+13+Revised+2022+(TR+13)+Fundamentals+of+an+Environmental+Monitoring+Program.">PDA Technical Report No. 13 Revised 2022 (TR 13) Fundamentals of an Environmental Monitoring Program.</a> Parenteral Drug Association, Inc. , (2022).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=USP&lt;1116&gt;+Microbiological+Evaluation+of+Cleanrooms+and+Other+Controlled+Environments+in+USP43-NF38+2S.">USP&lt;1116&gt; Microbiological Evaluation of Cleanrooms and Other Controlled Environments in USP43-NF38 2S.</a> United States Pharmacopeia - National Formulary. , (2020).</li><li>Guinet, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multicenter+study+on+incubation+conditions+for+environmental+monitoring+and+aseptic+process+simulation.">Multicenter study on incubation conditions for environmental monitoring and aseptic process simulation.</a> PDA journal of pharmaceutical science and technology. 71 (1), 43-49 (2017).</li><li>Gordon, O., Berchtold, M., Staerk, A., Roesti, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+different+incubation+conditions+for+microbiological+environmental+monitoring.">Comparison of different incubation conditions for microbiological environmental monitoring.</a> PDA Journal of Pharmaceutical Science and Technology. 68 (5), 394-406 (2014).</li><li>Anders, H. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multisite+qualification+of+an+automated+incubator+and+colony+counter+for+environmental+and+bioburden+applications+in+pharmaceutical+microbiology.">Multisite qualification of an automated incubator and colony counter for environmental and bioburden applications in pharmaceutical microbiology.</a> PDA Journal of Pharmaceutical Science and Technology. , (2022).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=USP&lt;1072&gt;+Disinfectants+and+Antiseptics.">USP&lt;1072&gt; Disinfectants and Antiseptics.</a> United States Pharmacopeia - National Formulary. , (2020).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=USP&lt;1071&gt;+Rapid+Microbial+Tests+for+Release+of+Sterile+Short-Life+Products:+A+Risk-Based+Approach+in+USP43-NF38+2S.">USP&lt;1071&gt; Rapid Microbial Tests for Release of Sterile Short-Life Products: A Risk-Based Approach in USP43-NF38 2S.</a> United States Pharmacopeia - National Formulary. , (2020).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PUSP&lt;1223&gt;+Validation+of+Alternative+Microbiological+Methods+in+USP43-NF38+2S.">PUSP&lt;1223&gt; Validation of Alternative Microbiological Methods in USP43-NF38 2S.</a> United States Pharmacopeia - National Formulary. , (2020).</li><li>Parenteral Drug Association. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PDA+Technical+Report+No.+33,+Revised+2013+(TR+33)+Evaluation,+Validation+and+Implementation+of+Alternative+and+Rapid+Microbiological+Methods.">PDA Technical Report No. 33, Revised 2013 (TR 33) Evaluation, Validation and Implementation of Alternative and Rapid Microbiological Methods.</a> Parenteral Drug Association, Inc. , (2013).</li><li>Putnam, N. E., Lau, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+study+identifies+a+sensitive,+low-risk,+closed-system+model+for+detection+of+fungal+contaminants+in+cell+and+gene+therapy+products.">Comprehensive study identifies a sensitive, low-risk, closed-system model for detection of fungal contaminants in cell and gene therapy products.</a> Journal of Clinical Microbiology. 59 (11), 0135721 (2021).</li><li>Pharmaceutical Inspection Convention &amp; Pharmaceutical Inspection Co-operation. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guide+to+Good+Manufacturing+Practice+for+Medicinal+Products+Annexes+(PE++009-16+(Annexes)).">Guide to Good Manufacturing Practice for Medicinal Products Annexes (PE 009-16 (Annexes)).</a> Pharmaceutical Inspection Convention &amp; Pharmaceutical Inspection Co-operation Scheme. , (2022).</li><li>United States Pharmacopeia - National Formulary. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=USP&lt;825&gt;+Radiopharmaceuticals+-+Preparation,+Compounding,+Dispensing,+and+Repackaging+in+USP43-NF38+2S.">USP&lt;825&gt; Radiopharmaceuticals - Preparation, Compounding, Dispensing, and Repackaging in USP43-NF38 2S.</a> United States Pharmacopeia - National Formulary. , (2020).</li></ol>

Yazarlar çıkar çatışması olmadığını beyan ederler.

Bu protokolde, temiz odalarda ve BSC'lerde aseptik tekniğin ve tek yönlü hava akışının sürdürülmesi de dahil olmak üzere birkaç kritik alan vardır. Aseptik manipülasyonlar, yukarıdan değil, ürünün yanından yapılmalıdır. Kapalı sistem işleme ve terminal olarak sterilize edilmiş hammaddelerin kullanılması tavsiye edilir. Kritik alanlarda konuşmaktan ve duvarlara veya ekipmanlara yaslanmaktan kaçınılmalıdır. Benzer şekilde, steril olmayan eşyalara gereksiz yere dokunmaktan ve düşen eşy...

Çevresel izleme (EM), proses izleme ve ürün sterilite testi yoluyla güçlü bir mikrobiyal izleme programının uygulanması, hücresel terapi laboratuvarlarında mevcut iyi doku uygulamaları (cGTP) ve mevcut iyi üretim uygulamaları (cGMP) için düzenleyici bir gerekliliktir1. Ek olarak, Amerika Birleşik Devletleri Gıda ve İlaç Dairesi (FDA), ürünün kalite kontrol (QC) testini gerçekleştiren laboratuvarın da aseptik dolum işlemleri için kullanılanlarla karşılaştırılabilir tesisler ve kontroller kullanmasını beklemektedir2. 
Bu protokolün dört ana bölümü vardır: 1) Personel önlükleme, temizlik ve malzemelerin sahnelenmesi dahil olmak üzere aseptik uygulamalar; 2) EM, uygulanabilir hava ve yüzey kültürleri ve canlı olmayan partikül hava izleme dahil; 3) çökeltme plakaları ve eldivenli parmak ucu örneklemesi dahil olmak üzere proses izleme; ve 4) özet Amerika Birleşik Devletleri Farmakopesi (USP) <71> yöntem3 veya NIH Alternatif Sterilite Test Yöntemi4 aracılığıyla ürün sterilite testi. Birlikte kullanıldığında, bu önlemler bir tesisin kontrol durumunda kalmasını sağlamak için etkili bir yöntem olabilir.
Burada anlatılan teknikler yeni değildir; Bununla birlikte, düzenleyicilerden ve meslek örgütlerinden gelen mevcut standartlar ayrıntıdan yoksundur, bu da özellikle yerinde üretim ve ürün sterilite testlerinin hızlı bir şekilde ortaya çıktığı akademik merkezlerde, mikrobiyal izlemenin veya standart dışı uygulamaların uygulanmasının olmamasına neden olmuştur 1,5,6 . Bu protokol, son kullanıcı doğrulama ve risk değerlendirmeleri ile birlikte kullanıldığında yasal gereklilikleri karşılayan bir mikrobiyal izleme ve kontrol programı oluşturmak için bir kılavuz olarak kullanılabilir.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>20-25&#176;C Incubator</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>30-35&#176;C Incubator</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Alcohol-based hand sanitizer</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>BacT/ALERT Dual-T instrument</td>
			<td>BioMerieux Industry</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Beard cover</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Biosafety cabinet (BSC)</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Cleanroom shoes</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Fluidthioglycollate medium (FTM)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>U84</td>
			<td>USP</td>
		</tr>
		<tr>
			<td>Handheld cleaning mop</td>
			<td>Contec</td>
			<td>2665LF</td>
			<td></td>
		</tr>
		<tr>
			<td>Hypodermic needle</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>iFA+ BacT/ALERT bottle</td>
			<td>Biomerieux</td>
			<td>412990</td>
			<td></td>
		</tr>
		<tr>
			<td>iFN+ BacT/ALERT bottle</td>
			<td>Biomerieux</td>
			<td>412991</td>
			<td></td>
		</tr>
		<tr>
			<td>Isokinetic head</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Laser particle counter</td>
			<td>TSI Incorporated</td>
			<td>9500-01</td>
			<td></td>
		</tr>
		<tr>
			<td>LpH III</td>
			<td>Steris</td>
			<td>1S16CX</td>
			<td></td>
		</tr>
		<tr>
			<td>Mirror</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Non-sterile bouffant</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Non-sterile gloves</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Non-sterile shoe covers</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Non-sterile sleeve covers</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Peridox RTU</td>
			<td>Contec</td>
			<td>CR85335IR</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic bag</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Sabouraud Dextrose Agar with Lecithinase and Tween (SABLT)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>P595</td>
			<td>USP, irradiated</td>
		</tr>
		<tr>
			<td>Sabouraurd Dextrose Agar (SAB)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>W565</td>
			<td>USP, irradiated</td>
		</tr>
		<tr>
			<td>Safety glasses</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Scrubs (top and bottom)</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Spor-Klenx RTU</td>
			<td>Steris</td>
			<td>6525M2</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile 70% isopropyl alcohol (IPA)</td>
			<td>Decon CiDehol</td>
			<td>8316</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile alcohol wipe</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Sterile boot covers</td>
			<td>Kimberly Clark</td>
			<td></td>
			<td>Cat# varies based on size</td>
		</tr>
		<tr>
			<td>Sterile coveralls</td>
			<td>Kimberly Clark</td>
			<td></td>
			<td>Cat# varies based on size</td>
		</tr>
		<tr>
			<td>Sterile face mask</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Sterile gloves</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Sterile hood</td>
			<td>Kimberly Clark</td>
			<td></td>
			<td>Cat# varies based on size</td>
		</tr>
		<tr>
			<td>Sterile low-lint wipes</td>
			<td>Texwipe</td>
			<td>TX3210</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile mop cleaning pads</td>
			<td>Contec</td>
			<td>MEQT0002SZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile sleeve covers</td>
			<td>Kimberly Clark</td>
			<td>36077</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile spreading rod</td>
			<td>Fisher Scientific</td>
			<td>14665231</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile syringe</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Tacky mats</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Tryptic Soy Agar (TSA)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>W570R</td>
			<td>USP, irradiated</td>
		</tr>
		<tr>
			<td>Tryptic Soy Agar with Lecithinase and Tween (TSALT)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>P520R</td>
			<td>USP, irradiated</td>
		</tr>
		<tr>
			<td>Tryptic Soy Broth (TSB)</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>U46</td>
			<td>USP</td>
		</tr>
		<tr>
			<td>Tubing</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
		<tr>
			<td>Vesphene III</td>
			<td>Steris</td>
			<td>1S15CX</td>
			<td></td>
		</tr>
		<tr>
			<td>Viable air sampler</td>
			<td>Hardy Diagnostics&#160;</td>
			<td>BAS22K</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex</td>
			<td></td>
			<td></td>
			<td>Lab preference</td>
		</tr>
	</tbody>
</table>

microbial control and monitoring strategies for cleanroom environments and cellular therapies

Sağlam önlük, temizlik, çevresel izleme ve personel izleme önlemlerini içeren iyi doğrulanmış ve bütünsel bir program, tesislerin kontrol durumunda çalışmasını sağlamak için hücresel terapi üretim paketlerinde ve ilgili test laboratuvarlarında mikrobiyal biyoyükü en aza indirmek için kritik öneme sahiptir. Sterilite testi gibi kalite kontrol önlemleri yoluyla ürün güvenliğinin sağlanması, hem minimum düzeyde manipüle edilmiş (bölüm 361) hem de minimum düzeyde manipüle edilmiş (bölüm 351) insan hücreleri, dokuları ve hücresel ve doku bazlı ürünler (HCT/P) için düzenleyici bir gerekliliktir. Bu videoda, Amerika Birleşik Devletleri Farmakopesi (USP<71>) ve Ulusal Sağlık Enstitüleri (NIH) Alternatif Sterilite Test Yöntemi tarafından sağlanan, önlük, temizlik, malzemelerin evrelendirilmesi, çevresel izleme, süreç izleme ve doğrudan aşılama kullanılarak ürün sterilite testi dahil olmak üzere temiz oda ortamında çalışmak için en iyi aseptik uygulamaların nasıl geliştirileceği ve dahil edileceği konusunda adım adım bir kılavuz sunuyoruz. Bu protokol, mevcut iyi doku uygulamalarını (cGTP) ve mevcut iyi üretim uygulamalarını (cGMP) karşılaması beklenen kuruluşlar için bir referans kılavuzu olarak tasarlanmıştır.

Implementing a strong microbial monitoring program through environmental monitoring (EM), process monitoring, and product sterility testing is a regulatory requirement for current good tissue practices (cGTP) and current good manufacturing practices (cGMP) in cellular therapy laboratories1. Additionally, the United States Food and Drug Administration (FDA) expects that the laboratory performing the quality control (QC) testing of the product should also employ facilities and controls comparable to those used for aseptic filling operations2. 
This protocol has four main sections: 1) Aseptic practices, including personnel gowning, cleaning, and staging of materials; 2) EM, including viable air and surface cultures and non-viable particle air monitoring; 3) process monitoring, including settling plates and gloved fingertip sampling; and 4) product sterility testing via the compendial United States Pharmacopeia (USP) &lt;71&gt; method3 or the NIH Alternative Sterility Testing Method4. When used together, these measures can be an effective method for ensuring that a facility remains in a state of control.
The techniques described here are not novel; however, current standards from regulators and professional organizations lack detail, which has led to an absence of microbial monitoring or the implementation of non-standardized practices, particularly in academic centers where on-site manufacturing and product sterility testing are emerging at a rapid rate1,5,6. This protocol can be used as a guide to create a microbial monitoring and control program that meets regulatory requirements when used in conjunction with end-user validation and risk assessments.

Yazar Spotlight: Temiz Oda Ortamları ve Hücresel Tedaviler için Mikrobiyal Kontrol ve İzleme Stratejileri  

Temiz Oda Ortamları ve Hücresel Terapiler için Mikrobiyal Kontrol ve İzleme Stratejileri

Clinical microbiology labs are being increasingly asked to assist with sterility testing as cellular therapies emerge as frontline treatment options. This project aims to provide clear instructions for implementing microbial monitoring and testing strategies applicable to any CGMP setting with a particular focus on academic and clinical laboratories. Gold standard microbial testing and CGMP relies on traditional culture-based methods, which are slow.While newer technologies exist, implementation is often limited by the regulatory requirement to conduct rigorous end-user validations to demonstrate non-inferiority. We hope that future work will help establish newer technologies into the mainstream for CGMP testing. There is no standardization between programs in CGMP.While a regulatory framework is provided, microbial monitoring and testing strategies are unique to each facility and rely on a risk-based approach and end-user validation. This can be confusing for establishing new CGMP programs in the clinical setting. Our group aims to bring awareness of CGMP to the clinical setting.We have published large-scale studies highlighting the advantages and pitfalls of blood culture systems for sterility testing as well as review articles demonstrating major differences between the clinical and CGMP microbiology labs and areas for consideration if implementing sterility testing in the clinical setting.

Beklenen sonuçlar Tablo 1'de açıklanmıştır. EM verileri gözden geçirilmeli ve uygun bir araştırma ve eylem, uyarı veya ISO limit sapmalarına yanıt verilerek takip edilmelidir. Canlı olmayan parçacıklar için bir sapma meydana gelirse, ISO 14644-Ek A, saniye A.5.57'ye göre devam edilmelidir. Gezi hemen tanımlanabilir anormal bir olaya atfedilebiliyorsa, orijinal örnekleme sonuçları belgelenmeli, orijinal sonuçları göz ardı etmek için bir not eklenmeli ve a...

Watch this Scientific Journal Video about Microbial Control and Monitoring Strategies for Cleanroom Environments and Cellular Therapies at JoVE.com

Protokol, temiz oda ortamında mikrobiyal biyolojik yükü en aza indirmek için en iyi uygulamaları özetler ve çevresel izleme, proses izleme ve ürün sterilite testi gibi stratejileri içerir. Mevcut iyi doku uygulamaları standartlarını ve mevcut iyi üretim uygulamaları standartlarını karşılamak için gerekli olan üretim ve test tesisleri ile ilgilidir.

A well-validated and holistic program that incorporates robust gowning, cleaning, environmental monitoring, and personnel monitoring measures is critical ...

Klinik mikrobiyoloji laboratuvarlarından, hücresel tedaviler ön saflarda tedavi seçenekleri olarak ortaya çıktıkça, sterilite testine yardımcı olmaları giderek daha fazla istenmektedir. Bu proje, özellikle akademik ve klinik laboratuvarlara odaklanarak herhangi bir CGMP ortamına uygulanabilir mikrobiyal izleme ve test stratejilerinin uygulanması için net talimatlar sağlamayı amaçlamaktadır. Altın standart mikrobiyal test ve CGMP, yavaş olan geleneksel kültür tabanlı yöntemlere dayanır.Daha yeni teknolojiler mevcut olsa da, uygulama genellikle aşağılık olmadığını göstermek için titiz son kullanıcı doğrulamaları yapmak için düzenleyici gereklilikle sınırlıdır. Gelecekteki çalışmaların CGMP testi için yeni teknolojilerin ana akım haline getirilmesine yardımcı olacağını umuyoruz. CGMP'de programlar arasında standardizasyon yoktur.Düzenleyici bir çerçeve sağlanırken, mikrobiyal izleme ve test stratejileri her tesise özgüdür ve riske dayalı bir yaklaşıma ve son kullanıcı doğrulamasına dayanır. Bu, klinik ortamda yeni CGMP programları oluşturmak için kafa karıştırıcı olabilir. Grubumuz CGMP farkındalığını klinik ortama getirmeyi amaçlamaktadır.Sterilite testi için kan kültürü sistemlerinin avantajlarını ve tuzaklarını vurgulayan büyük ölçekli çalışmaların yanı sıra, klinik ve CGMP mikrobiyoloji laboratuvarları arasındaki önemli farklılıkları ve klinik ortamda sterilite testinin uygulanmasında dikkate alınması gereken alanları gösteren makaleleri gözden geçirdik.

microbial-control-monitoring-strategies-for-cleanroom-environments

Research

JoVE Journal

Bioengineering

Microbial Control and Monitoring Strategies for Cleanroom Environments and Cellular Therapies

2.3K Görüntüleme.  National Institutes of Health. Klinik mikrobiyoloji laboratuvarlarından, hücresel tedaviler ön saflarda tedavi seçenekleri olarak ortaya çıktıkça, sterilite testine yardımcı olmaları giderek daha fazla istenmektedir. Bu proje, özellikle akademik ve klinik laboratuvarlara odaklanarak herhangi bir CGMP ortamına uygulanabilir mikrobiyal izleme ve test stratejilerinin uygulanması için net talimatlar sağlamayı amaçlamaktadır. Altın standart mikrobiyal test ve CGMP, yavaş olan geleneksel kültür tabanlı y?...

Video: Yazar Spotlight: Temiz Oda Ortamları ve Hücresel Tedaviler için Mikrobiyal Kontrol ve İzleme Stratejileri  

A well-validated and holistic program that incorporates robust gowning, cleaning, environmental monitoring, and personnel monitoring measures is critical for minimizing the microbial bioburden in cellular therapy manufacturing suites and the corresponding testing laboratories to ensure that the facilities are operating in a state of control. Ensuring product safety via quality control measures, such as sterility testing, is a regulatory requirement for both minimally manipulated (section 361) and more than minimally manipulated (section 351) human cells, tissues, and cellular and tissue-based products (HCT/Ps). In this video, we provide a stepwise guide for how to develop and incorporate the best aseptic practices for operating in a cleanroom environment, including gowning, cleaning, staging of materials, environmental monitoring, process monitoring, and product sterility testing using direct inoculation, provided by the United States Pharmacopeia (USP&lt;71&gt;) and the National Institutes of Health (NIH) Alternative Sterility Testing Method. This protocol is intended as a reference guide for establishments expected to meet current good tissue practices (cGTP) and current good manufacturing practices (cGMP).

The protocol summarizes the best practices to minimize microbial bioburden in a cleanroom environment and includes strategies such as environmental monitoring, process monitoring, and product sterility testing. It is relevant for manufacturing and testing facilities that are required to meet current good tissue practice standards and current good manufacturing practice standards.

Biyomühendislik

Gowning for a Cleanroom Space and Donning for a Biological Safety Cabinet

To begin, change into long-sleeved scrub top or short-sleeved scrub top with non-sterile sleeves, full-length scrub pants, and cleanroom shoes with shoe covers on. Sanitize hands with an alcohol-based hand sanitizer, and don non-sterile gloves, a beard cover if applicable, and a non-sterile bouffant. Remove the shoe covers, and step onto the tacky mat in front of the International Organization for Standardization or ISO-8 anteroom door.After discarding the shoe covers, decontaminate the gloves and door handle with 70%sterile isopropyl alcohol before entering the ISO 8 anteroom. Gather a sterile hood, face mask, coveralls, boot covers, and safety glasses. After decontaminating the gloves again, don safety glasses and stage the materials onto a clean surface.Don a new pair of non-sterile shoe covers over the dedicated cleanroom shoes, step onto the tacky mat, and enter the secondary ISO 7 anteroom, remaining on the dirty side of the demarcation line. Don the sterile hood and the sterile coverall, ensuring the neck of the hood is tucked inside the coverall to create a complete seal. Don the secondary sterile boot covers and decontaminate the gloves between each donning step before entering the ISO 7 space.To don into the biological safety cabinet, don sterile sleeves over the coverall, using thumb loops if present. Don sterile gloves over the existing gloves, ensuring the cuff of the glove extends over the sterile sleeve before entering the biological safety cabinet.

Temiz oda alanı için önlük giyme ve biyolojik güvenlik dolabı giyme

Settling Plates, Gloved Fingertip Sampling, and Sterility Testing by Direct Product Inoculation

Place the labeled TSA and SAB settling plates near the work area but in an area that will not interfere with testing. Document the sampling start time. Aseptically, open both plates, placing the lids on a clean surface in the biological safety cabinet away from the work area for up to four hours to prevent the agar from drying out.At the end of the processing close the lids of the plates and document the sampling stop time. Loosely, bag the plates and incubate the cultures. For gloved fingertip sampling, obtain two labeled TSALT plates.Aseptically remove the lid of the plates and place it on a clean work surface away from the sampling area. Gently roll the pad of each finger and thumb on the surface of the plate. Use sufficient force to make slight depressions on the agar without cracking the agar surface.Aseptically place the lid back onto the plate. After sanitizing the hands at the completion of sampling, loosely bag the plates and incubate the cultures. For direct inoculation, following the United States Pharmacopeia method, place the TSA and SAB settling plates in the biosafety cabinet.After preparing the cabinet and staging the materials, document the sampling start time. Obtain the test article and the associated tryptic soy broth and fluid thioglycollate medium bottles. If the bottles have a septum, remove the protective caps from each bottle and wipe the septum with a sterile alcohol wipe.Vortex the test article to ensure a homogenous suspension and an inoculate the bottles with a validated volume of the test article using a sterile syringe and a hypodermic needle. Wipe the septum with a sterile alcohol wipe and insert a sterile venting unit to allow for air exchange during incubation. Repeat the steps for each test article of that session.Aseptically close the TSA and SAB settling plates and document the sampling end time before performing the gloved fingertip sampling. Wipe the gloves with 70%sterile isopropyl alcohol, loosely bag the plates and incubate the cultures. Transfer all the test materials out of the biological safety cabinet.Similarly, for direct inoculation, following the NIH alternative sterility testing method, place the TSA and SAB settling plates in the biosafety cabinet and document the sampling start time. Obtain the test article and the associated IFA Plus labeled, IFN Plus labeled and SAB plate. Remove the protective caps from the IFA Plus and IFN Plus bottles and wipe the septum with a sterile alcohol wipe.After vortexing the test article, inoculate the bottles with validated volume of the test article using a sterile syringe and a hypodermic needle and wipe the septum again. Incubate the bottles on the BACT/ALERT Dual T instrument. Using a pipetter, inoculate the SAB plate with a validated amount of the test article and streak for isolation using a sterile loop.Let stand for 10 minutes to ensure the sample does not run onto the plate lid when inverted for incubation Place each SAB plate inoculated with a test article into a plastic bag. Tie them loosely. Aseptically close the TSA and SAB settling plates.Document the sampling end time on the plates, and perform the gloved fingertip sampling. After sampling, wipe the gloves with 70%alcohol. Loosely bag the plates after wiping the gloves with alcohol and incubate the cultures.A TSA air settling plate illustrating a single colony of a contaminant cultured during passive air process monitoring in the biological safety cabinet is shown here. The TSALT surface sampling plate with three colony forming units of two distinct colony morphologies is shown in this figure. The TSALT surface culture with a single colony on the plate's edge indicates poor aseptic handling during the sampling process.The correct method for obtaining gloved fingertip samples using the largest surface area of each finger or thumb is shown in this image. The incorrect process where only the fingertip is sampled is shown here. An example of mold balls visible to the naked eye that failed to be automatically detected by the BACT/ALERT system is shown here.Based on these findings, terminal visual inspection of all BACT/ALERT bottles and the addition of the SAB plate for fungal culturing using the NIH alternative sterility testing method is recommended.