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Hygiene and RNAse-free working
Andreas Jefimowicz (*1984, Karlsruhe)
Training/employment industrial office clerk
Hv-Engineering
Training biotechnological laboratory assistant
Johannes-Gutenberg-School
Employment biotechnician, Celonic GmbH
Study of biotechnology,
Mannheim University of Applied Sciences
Employment Bio.Kitchen UnternehmerTUM
Clean room, Celonic GmbH
Production of antibodies for
immune cancer therapy
Antibodies are injected into
patients
Must be free of contaminants
Introduction
● handling personal protective equipment
● cleaning and disinfection
● prevent contaminations
● dispose dangerous waste
Some definitions…
Hygeia was the Greek goddess of
health, cleanliness and sanitation
the degree of purity correlates with health
Disinfection: Is the process of using a chemical reagent to significantly
reduce the concentration of a pathogenic agent (bacteria, viruses,
fungi, parasites)
Sterility: Is the state of being free of disease-causing
microorganisms
Contamination: is the present of a constituent,
impurity or some other undesirable element that spoils,
corrupts, infects, makes unfit or makes inferior a material,
physical body, natural environment, workplace etc.
Why do we need all this?
You need to protect yourself...
It is assumed that every person you are testing could
be infected with SARS-CoV-2
It is better to assume someone is infected and he or
she is not than the other way around
SARS-CoV-2 stands for “severe acute respiratory
syndrome coronavirus type 2”
Infection with the new coronavirus can
lead to symptoms such as cough,
runny nose, sore throat and fever,
sometimes also diarrhoea, breathing
problems and pneumonia
The main route of transmission is
droplet infection. This can occur
directly via the mucous membranes of
the respiratory tract or indirectly via
hands, which then come into contact
with the oral or nasal mucosa and the
conjunctiva.
You also need to protect the samples you are
working with from yourself…
Micrococcus is a airborne microorganism. It occurs ubiquitous
(everywhere)
Also when you breathe, you spread many microorganisms like
bacteria, fungi, slime molds and viruses
When detecting SARS-CoV-2 we have to
amplify the RNA of the virus
RNA is single stranded nucleic acids
RNAses are enzymes that lead to degradation of RNA
Sampling
room
Hygiene concept regarding a test centre
Sample
preparation
room
Reagent
preparation
room Amplification
room
The whole process g
high virus concentrat
high levels of possibl
contamination by am
Implementation of the mobile laboratory in premises such as a hospital
Ideally entry to the laboratory rooms is
via a seal / lock room
Equipment is passed through
material locks
What do we need?
● personal protective equipment (PPE): overall, overshoes,
goggles, ffp2 mask, tape, gloves, lab coat
● dismozon/khorsolin/bleach
● rag
● sprayer
● wash bottle
● RNAse-Erase
● water
● autoclave
● wastebags
● Ethanol 70 %
● soap
● disinfectant
moisten hands
Hand Hygiene - Washing Hands (40-60 sec)
apply sufficient soap rub palms in circular motion rub palms and back of hands
Palms on top of each other
and fingers interlocked for
clean finger spaces
cleaning the back of
the fingers
Hold your thumb
and clean with a
twisting motion
Rub the fingertips on the
palm of each hand
Wash off soap with
water
Carefully dry with
disposable towels
Turn off the tap using
a disposable towel
Your hands are clean now
Gloves are important
- as a additional barrier
- to protect your skin
- to protect you from the virus
- to protect samples from you
wear them all the time, change or disinfect them
between steps in the protocol
Bleaches...
● have broad spectrum bactericidal properties
making them useful for disinfecting and sterilizing
● are able to inactivate bacteria, viruses, and algae
by oxidizing them
● chlorine bleach is a strong oxidizing agent,
so you need to protect you skin and eyes
RNase ERASE® ...
is a precisely formulated
reagent, ready-for-use,
ribonuclease
decontaminant
We need to use it when
working with RNA
to prevent degradation
What are the do´s and dont´s in a lab ?
Do´s
● When using the lab the first time clean with Dismozon, water and ethanol
● wash and disinfect your hands before and after working, tie back long hair
● change gloves and disinfect gloves with ethanol
● disinfect patient sampling room with ethanol between every patient
● treat working surfaces with RNAse-Erase when working with RNA
● wear an overall in the patient sampling room and the sample preparation room
● wear a labcoat in reagent preparation room and amplification room
● only touch what you have to touch while working
● physical distancing of at least 1.5 meter when working together
Dont´s
● do not touch your face or hair while working,
● do not touch the lower part of your pipette
● avoid coughing or sneezing, and don´t talk too much
● absolutely avoid touching non-sterile items and surfaces
● every morning wash hands and put on PPE, prepare Khorsolin, clean
handhelds, door knobs, surfaces, - wait 15 minutes, repeat with water
● wipe surfaces and fronts with ethanol and let them dry
● prepare your work area before you start
● in the patient treatment room use ethanol between patients to disinfect
● in the RNA Extraction area wipe surfaces/materials with RNAse-Erase
and repeat with ethanol
● same in RT-LAMP and qPCR area
● while handling patient samples and positive control wipe working area and
change gloves afterwards
● in assay run area clean with RNAse-Erase and ethanol, clean lids
● every evening clean floor with Khorsolin, wait 15 minutes, again clean
floor with water
Time Schedule
Sources:
Clean room: https://www.indiamart.com/proddetail/clean-room-for-pharmaceuticals-8688864812.html
Hygeia: https://www.flickr.com/photos/oar/16103293053
Bath house: https://www.pinterest.de/pin/72972456435506089/
Disinfectant agent: https://www.praxisdienst.de/Hygiene/Desinfektionsmittel/
Virus: https://pink.pharmaintelligence.informa.com/PS124172/USP-Sparks-Debate-Over-Rapid-Sterility-
Testing-For-Cell-And-Gene-Therapies
Biohazard: https://de.freepik.com/fotos-vektoren-kostenlos/biohazard-symbol
discussion round:
https://thenestline.com/staff_name/mahee-bhatt/
droplet infection: https://de.wikipedia.org/wiki/Tr%C3%B6pfcheninfektion
pneumonia: https://news.northwestern.edu/stories/2021/01/why-covid-19-pneumonia-worse-than-typical-
pneumonia/
micrococcus: https://www.istockphoto.com/de/foto/kolonien-von-micrococcus-luteus-bakterien-auf-agar-
platte-gm1148144414-309962622
airborne collector: https://www.holbach.biz/de/seite_LKS-30-amp-LKS100_15_mikrobiologie.html
RNA-virus: https://www.scinexx.de/dossierartikel/was-sind-viren/
RNA-DNA: https://www.drze.de/im-blickpunkt/abbildungen/DNA_RNA_2-de%20copyrighted.jpg/view

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Hygiene Concept.pdf

  • 2. Andreas Jefimowicz (*1984, Karlsruhe) Training/employment industrial office clerk Hv-Engineering Training biotechnological laboratory assistant Johannes-Gutenberg-School Employment biotechnician, Celonic GmbH Study of biotechnology, Mannheim University of Applied Sciences Employment Bio.Kitchen UnternehmerTUM
  • 3. Clean room, Celonic GmbH Production of antibodies for immune cancer therapy Antibodies are injected into patients Must be free of contaminants
  • 4.
  • 5.
  • 6.
  • 7. Introduction ● handling personal protective equipment ● cleaning and disinfection ● prevent contaminations ● dispose dangerous waste
  • 8. Some definitions… Hygeia was the Greek goddess of health, cleanliness and sanitation the degree of purity correlates with health
  • 9. Disinfection: Is the process of using a chemical reagent to significantly reduce the concentration of a pathogenic agent (bacteria, viruses, fungi, parasites) Sterility: Is the state of being free of disease-causing microorganisms Contamination: is the present of a constituent, impurity or some other undesirable element that spoils, corrupts, infects, makes unfit or makes inferior a material, physical body, natural environment, workplace etc.
  • 10. Why do we need all this?
  • 11. You need to protect yourself... It is assumed that every person you are testing could be infected with SARS-CoV-2 It is better to assume someone is infected and he or she is not than the other way around SARS-CoV-2 stands for “severe acute respiratory syndrome coronavirus type 2”
  • 12. Infection with the new coronavirus can lead to symptoms such as cough, runny nose, sore throat and fever, sometimes also diarrhoea, breathing problems and pneumonia The main route of transmission is droplet infection. This can occur directly via the mucous membranes of the respiratory tract or indirectly via hands, which then come into contact with the oral or nasal mucosa and the conjunctiva.
  • 13. You also need to protect the samples you are working with from yourself… Micrococcus is a airborne microorganism. It occurs ubiquitous (everywhere) Also when you breathe, you spread many microorganisms like bacteria, fungi, slime molds and viruses
  • 14. When detecting SARS-CoV-2 we have to amplify the RNA of the virus RNA is single stranded nucleic acids RNAses are enzymes that lead to degradation of RNA
  • 15. Sampling room Hygiene concept regarding a test centre Sample preparation room Reagent preparation room Amplification room The whole process g high virus concentrat high levels of possibl contamination by am
  • 16. Implementation of the mobile laboratory in premises such as a hospital Ideally entry to the laboratory rooms is via a seal / lock room Equipment is passed through material locks
  • 17. What do we need? ● personal protective equipment (PPE): overall, overshoes, goggles, ffp2 mask, tape, gloves, lab coat ● dismozon/khorsolin/bleach ● rag ● sprayer ● wash bottle ● RNAse-Erase ● water ● autoclave ● wastebags ● Ethanol 70 % ● soap ● disinfectant
  • 18. moisten hands Hand Hygiene - Washing Hands (40-60 sec) apply sufficient soap rub palms in circular motion rub palms and back of hands Palms on top of each other and fingers interlocked for clean finger spaces cleaning the back of the fingers Hold your thumb and clean with a twisting motion Rub the fingertips on the palm of each hand Wash off soap with water Carefully dry with disposable towels Turn off the tap using a disposable towel Your hands are clean now
  • 19. Gloves are important - as a additional barrier - to protect your skin - to protect you from the virus - to protect samples from you wear them all the time, change or disinfect them between steps in the protocol
  • 20. Bleaches... ● have broad spectrum bactericidal properties making them useful for disinfecting and sterilizing ● are able to inactivate bacteria, viruses, and algae by oxidizing them ● chlorine bleach is a strong oxidizing agent, so you need to protect you skin and eyes
  • 21. RNase ERASE® ... is a precisely formulated reagent, ready-for-use, ribonuclease decontaminant We need to use it when working with RNA to prevent degradation
  • 22. What are the do´s and dont´s in a lab ?
  • 23. Do´s ● When using the lab the first time clean with Dismozon, water and ethanol ● wash and disinfect your hands before and after working, tie back long hair ● change gloves and disinfect gloves with ethanol ● disinfect patient sampling room with ethanol between every patient ● treat working surfaces with RNAse-Erase when working with RNA ● wear an overall in the patient sampling room and the sample preparation room ● wear a labcoat in reagent preparation room and amplification room ● only touch what you have to touch while working ● physical distancing of at least 1.5 meter when working together
  • 24. Dont´s ● do not touch your face or hair while working, ● do not touch the lower part of your pipette ● avoid coughing or sneezing, and don´t talk too much ● absolutely avoid touching non-sterile items and surfaces
  • 25. ● every morning wash hands and put on PPE, prepare Khorsolin, clean handhelds, door knobs, surfaces, - wait 15 minutes, repeat with water ● wipe surfaces and fronts with ethanol and let them dry ● prepare your work area before you start ● in the patient treatment room use ethanol between patients to disinfect ● in the RNA Extraction area wipe surfaces/materials with RNAse-Erase and repeat with ethanol ● same in RT-LAMP and qPCR area ● while handling patient samples and positive control wipe working area and change gloves afterwards ● in assay run area clean with RNAse-Erase and ethanol, clean lids ● every evening clean floor with Khorsolin, wait 15 minutes, again clean floor with water Time Schedule
  • 26. Sources: Clean room: https://www.indiamart.com/proddetail/clean-room-for-pharmaceuticals-8688864812.html Hygeia: https://www.flickr.com/photos/oar/16103293053 Bath house: https://www.pinterest.de/pin/72972456435506089/ Disinfectant agent: https://www.praxisdienst.de/Hygiene/Desinfektionsmittel/ Virus: https://pink.pharmaintelligence.informa.com/PS124172/USP-Sparks-Debate-Over-Rapid-Sterility- Testing-For-Cell-And-Gene-Therapies Biohazard: https://de.freepik.com/fotos-vektoren-kostenlos/biohazard-symbol discussion round: https://thenestline.com/staff_name/mahee-bhatt/ droplet infection: https://de.wikipedia.org/wiki/Tr%C3%B6pfcheninfektion pneumonia: https://news.northwestern.edu/stories/2021/01/why-covid-19-pneumonia-worse-than-typical- pneumonia/ micrococcus: https://www.istockphoto.com/de/foto/kolonien-von-micrococcus-luteus-bakterien-auf-agar- platte-gm1148144414-309962622 airborne collector: https://www.holbach.biz/de/seite_LKS-30-amp-LKS100_15_mikrobiologie.html RNA-virus: https://www.scinexx.de/dossierartikel/was-sind-viren/ RNA-DNA: https://www.drze.de/im-blickpunkt/abbildungen/DNA_RNA_2-de%20copyrighted.jpg/view