Name: standard operating procedure for lyssavirus surveillance of the bat population in taiwan
Uploaded: 2019-08-27T13:30:00
Description: Watch this Scientific Journal Video about Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan at JoVE.com

We thank Tien-Cheng Li, Yi-Tang Lin, Chia-Jung Tsai, and Ya-Lan Li for their assistance during this study. This study was supported by grant no. 107AS-8.7.1-BQ-B2 (1) from the Bureau of Animal and Plant Health Inspection and Quarantine, Council of Agriculture, Executive Yuan, Taiwan.

1. Safety precautions when handling lyssaviruses
<ol>
	<li>Ensure that all laboratory workers handling bat specimens receive pre-exposure rabies prophylaxis17. Monitor the rabies antibody levels of the workers beforehand and re-examine them every 6 months17. Follow-up rabies vaccination is required for those whose antibody levels are lower than 0.5 IU/mL17.</li>
	<li>Depending on the biosafety regulations of the country where the laboratory is loca...

<ol>
	<li>Kuzmin, I. V., Rupprecht, C. E., Nagarajan, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Basic+facts+about+lyssavirus.">Basic facts about lyssavirus.</a> Current laboratory techniques in rabies diagnosis, research, and prevention, volume 1. , 3-21 (2014).</li><li>Ar&#233;chiga Ceballos, N., 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=Novel+lyssavirus+in+bat,+Spain.">Novel lyssavirus in bat, Spain.</a> Emerging Infectious Diseases. 19 (5), 793-795 (2013).</li><li>Gunawardena, P. S., 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=Lyssavirus+in+Indian+Flying+Foxes,+Sri+Lanka.">Lyssavirus in Indian Flying Foxes, Sri Lanka.</a> Emerging Infectious Diseases. 22 (8), 1456-1459 (2016).</li><li>Hu, S. C., 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=Lyssavirus+in+Japanese+Pipistrelle,+Taiwan.">Lyssavirus in Japanese Pipistrelle, Taiwan.</a> Emerging Infectious Diseases. 24 (4), 782-785 (2018).</li><li>Nokireki, T., Tammiranta, N., Kokkonen, U. -. M., Kantala, T., Gadd, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tentative+novel+lyssavirus+in+a+bat+in+Finland.">Tentative novel lyssavirus in a bat in Finland.</a> Transboundary Emerging Diseases. 65 (3), 593-596 (2018).</li><li>Banyard, A. C., Evans, J. S., Luo, T. R., Fooks, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lyssaviruses+and+bats:+emergence+and+zoonotic+threat.">Lyssaviruses and bats: emergence and zoonotic threat.</a> Viruses. 6 (8), 2974-2990 (2014).</li><li>Pal, S. 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=Rabies+virus+infection+of+a+flying+fox+bat,+Pteropus+policephalus+in+Chandigarh,+Northern+India.">Rabies virus infection of a flying fox bat, Pteropus policephalus in Chandigarh, Northern India.</a> Tropical and Geographical Medicine. 32 (3), 265-267 (1980).</li><li>Smith, P. C., Lawhaswasdi, K., Vick, W. E., Stanton, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+rabies+virus+from+fruit+bats+in+Thailand.">Isolation of rabies virus from fruit bats in Thailand.</a> Nature. 216 (5113), 384 (1967).</li><li>Tang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pivotal+role+of+dogs+in+rabies+transmission,+China.">Pivotal role of dogs in rabies transmission, China.</a> Emerging Infectious Diseases. 11 (12), 1970-1972 (2005).</li><li>Kuzmin, I. V., 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=Bat+lyssaviruses+(Aravan+and+Khujand)+from+Central+Asia:+phylogenetic+relationships+according+to+N,+P+and+G+gene+sequences.">Bat lyssaviruses (Aravan and Khujand) from Central Asia: phylogenetic relationships according to N, P and G gene sequences.</a> Virus Research. 97 (2), 65-79 (2003).</li><li>Arguin, P. M., 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=Serologic+evidence+of+Lyssavirus+infections+among+bats,+the+Philippines.">Serologic evidence of Lyssavirus infections among bats, the Philippines.</a> Emerging Infectious Diseases. 8 (3), 258-262 (2002).</li><li>Lumlertdacha, B., 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=Survey+for+bat+lyssaviruses,+Thailand.">Survey for bat lyssaviruses, Thailand.</a> Emerging Infectious Diseases. 11 (2), 232-236 (2005).</li><li>Kuzmin, I. V., 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=Lyssavirus+surveillance+in+bats,+Bangladesh.">Lyssavirus surveillance in bats, Bangladesh.</a> Emerging Infectious Diseases. 12 (3), 486-488 (2006).</li><li>Reynes, J. -. M., 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=Serologic+evidence+of+lyssavirus+infection+in+bats,+Cambodia.">Serologic evidence of lyssavirus infection in bats, Cambodia.</a> Emerging Infectious Diseases. 10 (12), 2231-2234 (2004).</li><li>Nguyen, A. T., 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=Bat+lyssaviruses,+northern+Vietnam.">Bat lyssaviruses, northern Vietnam.</a> Emerging Infectious Diseases. 20 (1), 161-163 (2014).</li><li>Liu, Y., Zhang, S., Zhao, J., Zhang, F., Hu, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+Irkut+virus+from+a+Murina+leucogaster+bat+in+China.">Isolation of Irkut virus from a Murina leucogaster bat in China.</a> PLoS Neglected Tropical Diseases. 7 (3), 2097 (2013).</li><li>Kaplan, M. M., Meslin, F. X., Kaplan, M. M., Kiprowski, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+precautions+in+handling+rabies+virus.">Safety precautions in handling rabies virus.</a> Laboratory Techniques in Rabies, 4th Ed. , 3-8 (1996).</li><li>Smith, I., Wang, L. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bats+and+their+virome:+an+important+source+of+emerging+viruses+capable+of+infecting+humans.">Bats and their virome: an important source of emerging viruses capable of infecting humans.</a> Current Opinion in Virology. 3 (1), 84-91 (2013).</li><li>Corbet, G. B., Hill, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The mammals of the Indomalayan region: a systematic review. , (1992).</li><li>Mayer, F., von Helversen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cryptic+diversity+in+European+bats.">Cryptic diversity in European bats.</a> Proceedings of the Royal Society B: Biological Sciences. 268 (1478), 1825-1832 (2001).</li><li>Epstein, J. H., Field, H. E., Wang, L. F., Cowled, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anthropogenic+epidemics:+the+ecology+of+bat-borne+viruses+and+our+role+in+their+emergence.">Anthropogenic epidemics: the ecology of bat-borne viruses and our role in their emergence.</a> Bats and viruses: a new frontier of emerging infectious diseases. , 249-280 (2016).</li><li>. Protocol for postmortem diagnosis of rabies in animals by direct fluorescent antibody testing Available from: <a target="_blank" href="https://www.cdc.gov/rabies/pdf/rabiesdfaspv2.pdf">https://www.cdc.gov/rabies/pdf/rabiesdfaspv2.pdf</a> (2019)</li><li>Hayman, D. T. S., 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=A+universal+real-time+assay+for+the+detection+of+Lyssaviruses.">A universal real-time assay for the detection of Lyssaviruses.</a> Journal of Virological Methods. 177 (1), 87-93 (2011).</li><li>Franka, 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=A+new+phylogenetic+lineage+of+rabies+virus+associated+with+western+pipistrelle+bats+(Pipistrellus+hesperus).">A new phylogenetic lineage of rabies virus associated with western pipistrelle bats (Pipistrellus hesperus).</a> Journal of General Virology. 87 (8), 2309-2321 (2006).</li><li>Trimarchi, C. V., Smith, J. S., Press, A., Jackson, A. C., Wunner, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+evaluation.">Diagnostic evaluation.</a> Rabies, 1st ed. , 307-349 (2002).</li><li>Moldal, T., 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=First+detection+of+European+bat+lyssavirus+type+2+(EBLV-2)+in+Norway.">First detection of European bat lyssavirus type 2 (EBLV-2) in Norway.</a> BMC Veterinary Research. 13, 216 (2017).</li><li>Robardet, E., 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=Comparative+assay+of+fluorescent+antibody+test+results+among+twelve+European+National+Reference+Laboratories+using+various+anti-rabies+conjugates.">Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates.</a> Journal of Virological Methods. 191 (1), 88-94 (2013).</li><li>Hanlon, C. A., Nadin-Davis, S. A., Jackson, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+diagnosis+of+rabies.">Laboratory diagnosis of rabies.</a> Rabies, 3rd ed. , 409-459 (2013).</li><li>Fischer, M., 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=A+step+forward+in+molecular+diagnostics+of+lyssaviruses--results+of+a+ring+trial+among+European+laboratories.">A step forward in molecular diagnostics of lyssaviruses--results of a ring trial among European laboratories.</a> PLoS ONE. 8 (3), 58372 (2013).</li><li>David, D., 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=Rabies+virus+detection+by+RT-PCR+in+decomposed+naturally+infected+brains.">Rabies virus detection by RT-PCR in decomposed naturally infected brains.</a> Veterinary Microbiology. 87 (2), 111-118 (2002).</li><li>Robardet, E., Picard-Meyer, E., Andrieu, S., Servat, A., Cliquet, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=International+interlaboratory+trials+on+rabies+diagnosis:+an+overview+of+results+and+variation+in+reference+diagnosis+techniques+(fluorescent+antibody+test,+rabies+tissue+culture+infection+test,+mouse+inoculation+test)+and+molecular+biology+techniques.">International interlaboratory trials on rabies diagnosis: an overview of results and variation in reference diagnosis techniques (fluorescent antibody test, rabies tissue culture infection test, mouse inoculation test) and molecular biology techniques.</a> Journal of Virological Methods. 177 (1), 15-25 (2011).</li></ol>

This laboratory standard operating procedure (SOP) provides a serial process for testing bat samples for the presence of lyssavirus antigens in Taiwan. The key steps include the employment of FAT and RT-PCR. The selection of suitable samples and successful isolation of the virus are also important. Additionally, some troubleshooting was conducted during the monitoring of bat lyssaviruses. The major difference was the target animals. Initially (2008&#8211;2009), the target animals of bat lyssavirus surveillance were live ...

Viruses within the genus Lyssavirus are zoonotic pathogens. There are at least seven lyssavirus species associated with human cases1. In addition to the 16 species in this genus1,2,3, Taiwan bat lyssavirus (TWBLV)4 and Kotalahti bat lyssavirus5 have been recently identified in bats, but their taxonomic statuses have yet to be determined.
Bats are the natural hosts of most lyssaviruses, with the exception of Mokola lyssavirus and Ikoma lyssavirus, which have yet not been identified in any bats1,2,3,6. The information on lyssaviruses in Asian bats is still limited. Two uncharacterized lyssaviruses in Asian bats (one in India and the other in Thailand)7,8 have been reported. One human rabies case associated with a bat bite in China was reported in 2002, but the diagnosis was made only by clinical observation9. In Central Asia, Aravan lyssavirus was identified in the lesser mouse-eared bat (Myotis blythi) in Kyrgyzstan in 1991, and Khujand lyssavirus was identified in the whiskered bat (Myotis mystacinus) in Tajikistan in 200110. In South Asia, Gannoruwa bat lyssavirus was identified in the Indian flying fox (Pteropus medius) in Sri Lanka in 20153. In Southeast Asia, several serological studies on bats in the Philippines, Thailand, Bangladesh, Cambodia, and Vietnam showed variable seroprevalence11,12,13,14,15. Although Irkut lyssavirus was identified in the greater tube-nosed bat (Murina leucogaster) in Jilin Province, China in 201216, the exact species and locations of lyssaviruses in East Asian bat populations remain unknown.
To assess the presence of lyssavirus in Taiwanese bat populations, a surveillance program employing both direct FAT and RT-PCR was initiated. Taiwan bat lyssavirus was identified in two cases of the Japanese pipistrelle (Pipistrellus abramus)4 in 2016&#8211;2017. In the present article, a laboratory standard operating procedure is introduced for lyssavirus surveillance of the bat population in Taiwan. The flow chart of bat lyssavirus diagnosis in our laboratory is presented in Figure 1.

<table>
	<tbody>
		<tr>
			<td>2.5% Trypsin (10x)</td>
			<td>Gibco</td>
			<td>15090046</td>
			<td>Trppsin</td>
		</tr>
		<tr>
			<td>25 cm2 flask</td>
			<td>Greiner bio-one</td>
			<td>690160</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>Honeywell</td>
			<td>32201-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose I</td>
			<td>VWR Life Science</td>
			<td>97062-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol</td>
			<td>NIHON SHIYAKU REAGENT</td>
			<td>NS-32294</td>
			<td></td>
		</tr>
		<tr>
			<td>AMV Reverse Transcriptase</td>
			<td>Promega</td>
			<td>M5101</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic(100X)&#160;</td>
			<td>Gibco</td>
			<td>15240-062</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>Blade</td>
			<td>Braun</td>
			<td>BA215</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>eppendorf</td>
			<td>5424R</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemilumineance system</td>
			<td>TOP BIO CO.</td>
			<td>MGIS-21-C2-1M</td>
			<td></td>
		</tr>
		<tr>
			<td>Collection tube</td>
			<td>Qiagen</td>
			<td>990381</td>
			<td></td>
		</tr>
		<tr>
			<td>Collection tube</td>
			<td>SSI</td>
			<td>2341-SO</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover slide</td>
			<td>Muto Pure chemical Co., LTD.</td>
			<td>24505</td>
			<td></td>
		</tr>
		<tr>
			<td>DNA analyzer</td>
			<td>Applied Biosystems</td>
			<td>3700XL</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco</td>
			<td>10437028</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>FITC Anti-Rabies Monoclonal Globulin</td>
			<td>Fujirebio Diagnostic Inc.</td>
			<td>800-092</td>
			<td>FITC-conjugated anti-rabies antibodies: reagent B</td>
		</tr>
		<tr>
			<td>Four-well Teflon-coating glass slide</td>
			<td>Thermo Fisher Scientific</td>
			<td>30-86H-WHITE</td>
			<td></td>
		</tr>
		<tr>
			<td>Gel Electrophoresis System</td>
			<td>Major Science</td>
			<td>MJ-105-R</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS (1x)</td>
			<td>Gibco</td>
			<td>14175095</td>
			<td>Trppsin</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>ASTEC</td>
			<td>SCA-165DS</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted Microscope</td>
			<td>Olympus</td>
			<td>IX71</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Glutamine 200 mM (100x)</td>
			<td>Gibco</td>
			<td>A2916801</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>LIGHT DIAGNOSTICS Rabies FAT reagent</td>
			<td>EMD Millipore Corporation</td>
			<td>5100</td>
			<td>FITC-conjugated anti-rabies antibodies: reagent A</td>
		</tr>
		<tr>
			<td>MagNA Pure Compact Instrument</td>
			<td>Roche</td>
			<td>03731146001</td>
			<td></td>
		</tr>
		<tr>
			<td>MagNA Pure Compact NA Isolation Kit 1</td>
			<td>Roche</td>
			<td>03730964001</td>
			<td></td>
		</tr>
		<tr>
			<td>MEM (10x)</td>
			<td>Gibco</td>
			<td>11430030</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>MEM NEAA (100x)</td>
			<td>Gibco</td>
			<td>11140050</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>MEM vitamin solution</td>
			<td>Gibco</td>
			<td>11120052</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Merck</td>
			<td>1.06329.0500</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>Needle</td>
			<td>Terumo</td>
			<td>NN*2332R9</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Medicago</td>
			<td>09-8912-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Primer synthesis</td>
			<td>Mission Biotech</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>RNasin ribonuclease inhibitor</td>
			<td>Promega</td>
			<td>N2111</td>
			<td></td>
		</tr>
		<tr>
			<td>Sequencing service</td>
			<td>Mission Biotech</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Slide</td>
			<td>Thermo Scientific</td>
			<td>AA00008032E00MNT10</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Pyruvate (100 mM)</td>
			<td>Gibco</td>
			<td>11360070</td>
			<td>MEM-10</td>
		</tr>
		<tr>
			<td>Stainless Steel Beads</td>
			<td>QIAGEN</td>
			<td>69989</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile absorbent pad</td>
			<td>3M</td>
			<td>1604T-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe filter</td>
			<td>Nalgene</td>
			<td>171-0045</td>
			<td></td>
		</tr>
		<tr>
			<td>Taq polymerase</td>
			<td>JMR Holdings</td>
			<td>JMR-801</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermal cycler</td>
			<td>Applied Biosystems</td>
			<td>2720</td>
			<td></td>
		</tr>
		<tr>
			<td>TissueLyser II</td>
			<td>QIAGEN</td>
			<td>85300</td>
			<td></td>
		</tr>
		<tr>
			<td>Tongue depressor</td>
			<td>HONJER CO., LTD.</td>
			<td>122246</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezer</td>
			<td>Tennyson medical Instrument developing CO., LTD.</td>
			<td>A0601</td>
			<td></td>
		</tr>
		<tr>
			<td>Tylosin Tartrate</td>
			<td>Sigma</td>
			<td>T6271-10G</td>
			<td>MEM-10</td>
		</tr>
	</tbody>
</table>

standard operating procedure for lyssavirus surveillance of the bat population in taiwan

Viruses within the genus Lyssavirus are zoonotic pathogens, and at least seven lyssavirus species are associated with human cases. Because bats are natural reservoirs of most lyssaviruses, a lyssavirus surveillance program of bats has been conducted in Taiwan since 2008 to understand the ecology of these viruses in bats. In this program, non-governmental bat conservation organizations and local animal disease control centers cooperated to collect dead bats or bats dying of weakness or illness. Brain tissues of bats were obtained through necropsy and subjected to direct fluorescent antibody test (FAT) and reverse transcription polymerase chain reaction (RT-PCR) for detection of lyssavirus antigens and nucleic acids. For the FAT, at least two different rabies diagnosis conjugates are recommended. For the RT-PCR, two sets of primers (JW12/N165-146, N113F/N304R) are used to amplify a partial sequence of the lyssavirus nucleoprotein gene. This surveillance program monitors lyssaviruses and other zoonotic agents in bats. Taiwan bat lyssavirus is found in two cases of the Japanese pipistrelle (Pipistrellus abramus) in 2016&#8211;2017. These findings should inform the public, health professionals, and scientists of the potential risks of contacting bats and other wildlife.

From 2014 to May 2017, 332 bat carcasses from 13 species were collected for surveillance. Two tested positive. In the first bat case, the brain impression tested negative using the FAT with one of the commercial FITC-conjugated anti-rabies antibodies (Figure 2), while the RT-PCRs employing each of the two primer sets (JW12/N165-146, N113F/N304R) yielded positive results (Figure 3). A 428 bp sequence of amplicon (amplified with N113F/N304R and containing the part...

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Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan

This protocol introduces a standard laboratory operating procedure for diagnostic testing of lyssavirus antigens in bats in Taiwan.

Viruses within the genus Lyssavirus are zoonotic pathogens, and at least seven lyssavirus species are associated with human cases. Because bats are ...

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