(Tissue Transport SOP)

Guidelines for Tissue Removal, Transport, Manipulation, and Clean-up Procedures from MPV Positive and Suspect Areas to Outside Laboratories.

 

The University IACUC in conjunction with the DLAR has determined that all unfixed animal tissue, cells, or bodily fluids removed from confirmed positive or suspect rooms in the 10B corridor pose a significant risk of contamination of laboratory areas and equipment with the MPV virus. This virus is extremely resistant in the environment and can survive for weeks to months outside the host. Contaminated laboratory surfaces and equipment pose a potential for transfer of the virus to the clothing and skin of laboratory personnel; such surfaces, equipment, and personnel may then serve as fomites for re-infection of animal populations. For this reason, it is required that all unfixed tissues, cells, and bodily fluids removed from mice in confirmed positive or suspect housing areas be treated as potentially biohazardous material. To mitigate the risk of spread of MPV to future animal populations, all personnel working with such tissues must follow the following guidelines:

 

 

TRANSPORT OF MATERIAL FROM ANIMAL FACILTY TO LABORATORY

 

· All samples must be placed in a primary leak-proof container, which is then packed in a sealed leak-proof secondary container. Acceptable primary containers include petrie dishes, glass jars, or vacutainer tubes. Recommended secondary containers include ziplock plastic bags, or plastic jars or boxes with tops.

 

·The primary container must be packed in such a way that, under normal conditions of transport, it cannot break, be punctured or leak contents into the secondary container.

 

·When transporting liquids or tissues, there must be absorbent material (paper towel, cotton batting) placed between the primary receptacle and the secondary container. The absorbent material must be in sufficient quantity to absorb the entire liquid contents of the primary receptacle within the secondary container should the primary container be broken or leak.

 

·The outside surface of the secondary container must be thoroughly disinfected with either a 10% Clorox solution or chlorine based disinfectant such as Clidox prior to removal from the infected room or procedural use area. Clidox is available in the procedure rooms or from the facility staff or supervisors.

                                                                                        

·Sample must remain sealed while being transported to lab.

 

HANDLING OF MATERIAL WITHIN INVESTIGATOR LABORATORIES

 

·Lab must have restricted access at the time of sample manipulation and until effective disinfection has occurred subsequent to handling the material. Restricted access includes appropriate signage posted at the entrance to the laboratory to the effect that work with contaminated tissue, cells, or bodily fluids is occurring within. Persons who are not essential to the manipulation of such tissue should not enter the laboratory until disinfection has been completed.

 

·Prior to opening of secondary sealed container, full BSL2 garb must be worn inclusive of gloves, disposable Tyvek suit or disposable lab coat, and shoe covers.

 

·Sample must either be handled within a BSL2 hood or in a designated area within the lab that is free of clutter and can be adequately disinfected.

 

·All surface and contact areas, equipment and supplies (inclusive of pens and paperwork), must be thoroughly disinfected with either a 10% Clorox solution or chlorine based disinfectant such as Clidox after samples have been processed. Gas or steam autoclaving is also an acceptable method of decontamination for equipment.

 

·If sample manipulation includes procedures with the potential for aerosolization of virus particles, such as tissue homogenization, a consultation with EH&S is required. In addition to the above detailed disinfection techniques, chemical fogging of the laboratory area may also be required in these cases.

 

·All garb, as well as disposable material used in procedures (chucks, drapes, pipettes etc.) must be disposed of as biohazard waste. Such disposal includes double bagging of material, and disinfection of the exterior bag with Clorox or Clidox solution prior to moving it from the designated tissue handling area

 

·Discarded tissues must be bagged as above and treated as biohazardous waste.

 

USE OF CELL LINES FROM POSITIVE OR SUSPECT AREAS FOR CULTURE

 

·The use of tissues from potentially infected animals for cell culture is strongly discouraged. Such cultures present a risk of propagating the virus, thus maintaining a nidus of potential contamination and spread . Investigators are urged to utilize clean, non-infected animals as the source of such materials. While some acute in-vitro uses of such lines may not present a high risk of contamination, the propagation of long-term maintenance or “immortal” lines presents a significant risk. Cell lines intended for long-term maintenance in culture must be MAP or PCR tested for MPV prior to such use. Further information on such testing is available from the DLAR (648-8950).

 

TRANSFER OF CELLS, TISSUES, OR BODILY FLUIDS INTO LIVING ANIMALS

 

·Transfer of potentially infected cells, tissues, or bodily fluids into living animals is not permitted due to the potential of the spread of the virus to recipient animals. No such material may be transferred to other researchers for this purpose, nor may such materials be “banked” or frozen for future use in living animals.