Sampling Recommendations for Pet PCR Nucleic Acid Testing

Sample Type	Sampling Method	Pathogen
Eye, nose, and throat swab	Eye swab: moisten the swab head with sample preservation solution, and then gently wipe under the pet’s eye and conjunctiva 3-5 times; Tears can be appropriately collected as a sample. Nasal swab: collect by gently rubbing the tip of a disposable swab under the pet’s nasal cavity 3-5 times.Oral/pharyngeal swab: extend the disposable swab into the pet’s posterior pharyngeal wall and tonsillar crypts, gums swab 3-5 times to collect. Note: try to collect mucosal cells and avoid the collection of viscous secretions; Respiratory test can be used to swab the three parts of the sequential collection to improve the sampling effect.	FCV、FHV-1、Chl、Myc、CDV、CPIV、 IAV、CAV-2、Bb、etc.
Fecal samples Anal swab Intestinal lavage fluid	Fecal/anal swab: wet the swab head with SPB sample preservation solution, then insert it into the anus at about 1-2cm, gently rotate the swab, and collect mucus or feces on the surface of the colorectal tract ; After broking the swab into the SPB sample preservation solution, mix well and perform nucleic acid extraction; Note: if the sample is excessive, it must be further diluted and then extracted for nucleic acid extraction.	αCoV、FIP、 CPV、Cry、Gia、T.foetus、 Toxo、etc.
Blood	Take 200ul of EDTA anticoagulated whole blood, add to SPB sample preservation solution and mix well, then perform nucleic acid extraction. (Do not anticoagulate with heparin, do not centrifuge, and mix well if blood is stratified).	B. canis、Toxo、Lep、Ehr、 B. henselae、 H. myc、CAV-1、FIV、Felv、etc.
Pleural effusion Ascites Aqueous humor Lymph node aspirate	Pleural fluid/abdominal fluid: take 200ul of pleural fluid/abdominal fluid, add it to the SPB sample preservation solution, mix well and then take 200ul of liquid for nucleic acid extraction. (If the effusion sample is too thick or milky need to carry higher ratio of dilution) Aqueous humor, lymphatic puncture fluid, cerebrospinal fluid: 200ul of liquid can be taken directly for nucleic acid extraction; If the sample is less than 200ul, at least 50ul should be taken, and then make up 200 ul with SPB sample preservation solution and mix well. (If there are solid protein tissues in the lymphatic puncture fluid, proteinase K can be used to assist the dilution)	Feline Infectious Peritonitis（FIP）
Cerebrospinal fluid (CSF)	Take 200 ul of liquid directly for nucleic acid extraction; If less than 200 pl, take at least 50 ul or more, and then make up 200 ul with the sample preservation solution and then carry out nucleic acid extraction.	FIP、CDV、Toxo、B. henselae、etc.
Urine	Nucleic acid extraction was performed by directly taking 200 ul of urine.	Leptospira spp(Lep)
Microblood Feather	Prepare EDTA blood 0.1 mL or new feather (near chest and abdomen) 4-5 pieces or feces，Add 20ul proteinase k to the lysate, then add 20uled anticoagulated whole blood or 3-4 feathers to the lysate, mix well and wait for lysis for 15 minutes, then add 20ul magnetic bead suspension to the lysate and mix well, then add the whole lysate to the LB well for nucleic acid extraction. (The steps are special, you can ask the after-sales service for the operation instruction video)	Parrot Beak And Feather Disease VirusAvian Polyomavirus
Sample storage	It is recommended that nucleic acid extraction and testing should be performed immediately after sample collection; If storage is required, it can be stored at 4℃ for a short period of time or below -20℃ for a long period of time; When the samples are transported for testing, they need to be refrigerated for transportation.
Note	When different samples can be selected for testing, please select according to symptoms. When using swabs the swab tip should be quickly broken off into the SPB sample preservation solution after sampling. If the above sample extraction results are poor or infrequently used samples such as spinal fluid, genital secretions/abortive secretions、skin mucosal lesion swab. The criterion for determining the success of the extraction is whether there is any magnetic bead remaining in the LB wells after the completion of the extraction; If the magnetic bead residue is severe, the sample needs to be further diluted until there is no powder in the LB wells after the extraction.

Eye, nose, and throat swab

Eye swab: moisten the swab head with sample preservation solution, and then gently wipe under the pet’s eye and conjunctiva 3-5 times; Tears can be appropriately collected as a sample.

Nasal swab: collect by gently rubbing the tip of a disposable swab under the pet’s nasal cavity 3-5 times.Oral/pharyngeal swab: extend the disposable swab into the pet’s posterior pharyngeal wall and tonsillar crypts, gums swab 3-5 times to collect.

Note: try to collect mucosal cells and avoid the collection of viscous secretions; Respiratory test can be used to swab the three parts of the sequential collection to improve the sampling effect.

FCV、FHV-1、Chl、Myc、CDV、CPIV、
IAV、CAV-2、Bb、etc.

Fecal samples
Anal swab
Intestinal lavage fluid

Fecal/anal swab: wet the swab head with SPB sample preservation solution, then insert it into the anus at about 1-2cm, gently rotate the swab, and collect mucus or feces on the surface of the colorectal tract ;

After broking the swab into the SPB sample preservation solution, mix well and perform nucleic acid extraction;

Note: if the sample is excessive, it must be further diluted and then extracted for nucleic acid extraction.
PCR Nucleic Acid Testing

αCoV、FIP、 CPV、Cry、Gia、T.foetus、
Toxo、etc.

Blood

Take 200ul of EDTA anticoagulated whole blood, add to SPB sample preservation solution and mix well, then perform nucleic acid extraction. (Do not anticoagulate with heparin, do not centrifuge, and mix well if blood is stratified).

B. canis、Toxo、Lep、Ehr、 B. henselae、
H. myc、CAV-1、FIV、Felv、etc.

Pleural effusion
Ascites
Aqueous humor
Lymph node aspirate

Pleural fluid/abdominal fluid: take 200ul of pleural fluid/abdominal fluid, add it to the SPB sample preservation solution, mix well and then take 200ul of liquid for nucleic acid extraction. (If the effusion sample is too thick or milky need to carry higher ratio of dilution)

Aqueous humor, lymphatic puncture fluid, cerebrospinal fluid: 200ul of liquid can be taken directly for nucleic acid extraction; If the sample is less than 200ul, at least 50ul should be taken, and then make up 200 ul with SPB sample preservation solution and mix well. (If there are solid protein tissues in the lymphatic puncture fluid, proteinase K can be used to assist the dilution)

Feline Infectious Peritonitis（FIP）

Cerebrospinal fluid (CSF)

Take 200 ul of liquid directly for nucleic acid extraction; If less than 200 pl, take at least 50 ul or more, and then make up 200 ul with the sample preservation solution and then carry out nucleic acid extraction.

FIP、CDV、Toxo、B. henselae、etc.

Urine

Nucleic acid extraction was performed by directly taking 200 ul of urine.

Leptospira spp(Lep)

Microblood
Feather

Prepare EDTA blood 0.1 mL or new feather (near chest and abdomen) 4-5 pieces or feces，Add 20ul proteinase k to the lysate, then add 20uled anticoagulated whole blood or 3-4 feathers to the lysate, mix well and wait for lysis for 15 minutes, then add 20ul magnetic bead suspension to the lysate and mix well, then add the whole lysate to the LB well for nucleic acid extraction. (The steps are special, you can ask the after-sales service for the operation instruction video)

Parrot Beak And Feather Disease VirusAvian Polyomavirus

Sample storage

It is recommended that nucleic acid extraction and testing should be performed immediately after sample collection; If storage is required, it can be stored at 4℃ for a short period of time or below -20℃ for a long period of time; When the samples are transported for testing, they need to be refrigerated for transportation.

Note

When different samples can be selected for testing, please select according to symptoms. When using swabs the swab tip should be quickly broken off into the SPB sample preservation solution after sampling. If the above sample extraction results are poor or infrequently used samples such as spinal fluid, genital secretions/abortive secretions、skin mucosal lesion swab. The criterion for determining the success of the extraction is whether there is any magnetic bead remaining in the LB wells after the completion of the extraction; If the magnetic bead residue is severe, the sample needs to be further diluted until there is no powder in the LB wells after the extraction.