# KIs virus and blood donors, France.

Philippe Biagini, Mhammed Touinssi, Vital Galicher, Philippe de Micco

► To cite this version:

Philippe Biagini, Mhammed Touinssi, Vital Galicher, Philippe de Micco. KIs virus and blood donors, France. Emerging Infectious Diseases, 2012, 18 (8), pp.1374-1375. 10.3201/eid1808.120442. hal-00763185

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Submitted on 12 Oct 2023

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Pecuarias, Morelos, Mexico (Z. García); and Oklahoma State University, Stillwater, Oklahoma, USA (J. de la Fuente)

DOI: http://dx.doi.org/10.3201/eid1808.111902

References

1. Miller R, Kaneene JB, Fitzgerald SD, Schmitt SM. Evaluation of the influence of supplemental feeding of white-tailed deer (*Odocoileus virginianus*) on the prevalence of bovine tuberculosis in the Michigan wild deer population. J Wildl Dis. 2003;39:84-95.

2. Barrios-Garcia HB, Guizarnotegui-Blanco JA, Zapata-Campos CC, Almazán-García C, González-Alanís P, Villareal-Peña R, et al. Identification of *Mycobacterium tuberculosis* complex by histopathology and PCR in white-tailed deer (*Odocoileus virginianus*) in Tamaulipas, Mexico. Journal of Animal and Veterinary Advances. 2012;11:1036-40.

3. Pérez de León AA, Strickman DA, Knowles DP, Fish D, Thacker E, De La Fuente J, et al. One Health approach to identify research needs in bovine and human babesioses: workshop report. Parasit Vectors. 2010;3:36.

4. Pound JM, George JE, Kammlah DM, Lohmeyer KH, Davey RB. Evidence for role of white-tailed deer (Artiodactyla: Cervidae) in epizootiology of cattle ticks and southern cattle ticks (Acari: Ixodidae) in reinfestations along the Texas/Mexico border in south Texas: a review and update. J Econ Entomol. 2010;103:211-8. http://dx.doi.org/10.1603/EC09359

5. Lucero NE, Ayala SM, Escobar GJ, Jacob NR. *Brucella* isolated in humans and animals in Latin America from 1968 to 2006. Epidemiol Infect. 2008;136: 496-503. http://dx.doi.org/10.1017/S0950268807008795

6. Cooper K, Huang FF, Batista L, Rayo CD, Bezanilla JC, Toth TE, et al. Identification of genotype 3 hepatitis E virus (HEV) in serum and fecal samples from pigs in Thailand and Mexico, where genotype 1 and 2 HEV strains are prevalent in the respective human populations. J Clin Microbiol. 2005;43:1684-8. http://dx.doi.org/10.1128/JCM.43.4.1684-1688.2005

7. Boadella M, Casas M, Martín M, Vicente J, Segalés J, de la Fuente J, et al. Increasing contact with hepatitis E virus in red deer, Spain. Emerg Infect Dis. 2010;16:1994-6.

8. Cantú A, Ortega-S JA, Mosqueda J, García-Vazquez Z, Henke SE, George JE. Prevalence of infectious agents in free-ranging white-tailed deer in northeastern Mexico. J Wildl Dis. 2008;44:1002-7.

9. Boadella M, Lyashchenko K, Greenwald R, Esfandiari J, Jaroso R, Carta T, et al. Serologic tests for detecting antibodies against *Mycobacterium bovis* and *Mycobacterium avium* subspecies *paratuberculosis* in Eurasian wild boar (*Sus scrofa scrofa*). J Vet Diagn Invest. 2011;23:77-83. http://dx.doi.org/10.1177/104063871102300111

10. Boadella M, Barasona JA, Diaz-Sanchez S, Lyashchenko KP, Greenwald R, Esfandiari J, et al. Performance of immunochromatographic and ELISA tests for detecting fallow deer infected with *Mycobacterium bovis*. Prev Vet Med. 2012;104:160-4. http://dx.doi.org/10.1016/j.prevetmed.2011.10.005

Address for correspondence: Christian Gortazar, Instituto de Investigación en Recursos Cinegéticos, Ronda de Toledo s/n, 13005 Ciudad Real, Spain; email: christian.gortazar@uclm.es

KIs Virus and Blood Donors, France

**To the Editor:** KIs-V is a new putative virus identified recently in the blood of persons in Japan (1). First partial sequence of KIs-V was characterized unexpectedly, when PCR primers were used that were directed primarily to the consensus domain of helicase of positive-stranded RNA viruses. Extensive physicochemical and molecular analysis suggested that KIs-V is an enveloped virus with a circular, double-stranded DNA genome of ≈9,500 bp (prototype isolate: GenBank accession no. AB550431); its genetic diversity is presumed to be extremely low because the 4 complete genomes already characterized in Japan harbor strict identical sequences. The 13 potential genes identified by in silico analysis exhibit an overall low sequence homology to other known viral proteins (1). Until now, KIs-V epidemiologic data have been related

only to the original study in which the authors analyzed plasma samples from 516 blood donors categorized into 4 groups by alanine aminotransferase (ALT) level (either ≤60 IU/L or >60 IU/L) and the presence or absence of hepatitis E virus (HEV) antibodies. As a result, KIs-V DNA was detectable at elevated prevalence in the high ALT level/HEV antibody-positive group (36%, n = 100); viral loads, checked for a few samples, ranged from 10⁶ to 10⁸ copies/mL. KIs-V DNA also was identified in HEV antibody-negative samples, with low or high ALT level (≈0.8%, n = 120, and 1%, n = 100, respectively) (1).

To gain insights about the potential presence of this virus in the blood of persons in France, we investigated KIs-V DNA in the plasma of 576 healthy blood donors (mean age 40 years; 306 men; men:women 1:1.13). Blood samples were collected in vacuum tubes (Vacutainer, SST, Becton Dickinson, Meylan, France) and centrifuged, and plasma aliquots were stored at -80°C until use. Nucleic acids were extracted from 1-mL plasma volumes (MagNA Pure LC, Roche Diagnostics, Meylan, France) (2) and tested for KIs-V DNA by using the same nested PCR system for screening Japanese blood donors (1). Briefly, one tenth (5 µL) of extracted nucleic acids were first amplified by using primers 101-C (5'-CAACACCGCA ATCACAAAGT-3') and N101-B (5'-AACATTGAAACGTCATGT CC-3') (0.8 µM each) in a 50-µL mix containing deoxynucleotide triphosphates (0.2 mM each) (Roche) and 2 U Taq DNA polymerase (Invitrogen, Cergy Pontoise, France) with its corresponding buffer. One microliter of the amplification mixture was subsequently used in a second-round PCR with primers KS-2 (5'-CTCGTCCTCGTCGTCATC GTA-3') and N101-D (5'-CA TTTGCTCCCGCTGGAGATG-3') under the same conditions as above. The amplification conditions for first-
and second-round PCR were 94°C for 3 min, followed by 40 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 2 min. Expected amplification products were 458 bp (PCR-1) and 304 bp (PCR-2). Using dilutions of a synthetic template corresponding to the target sequence, we estimated the sensitivity of the amplification assay to be <5 copies of target sequence by limiting-dilution assay.

Negative (sterile water) and positive controls (synthetic template dilutions) were added systematically to each amplification run. A PCR control intended to check the quality of the nucleic acids extraction procedure was also performed systematically on 4 randomly selected samples of each batch (n = 32); this control was based on the detection of an extremely prevalent DNA virus (Torque Teno virus and related viruses, family *Anelloviridae*) by using a highly conserved amplification system (3).

Among the 576 plasma samples tested, no positive signal was identified for KIs-V DNA after agarose gel electrophoresis of PCR-1 and PCR-2. Amplification controls (negative, positive, anellovirus DNA) confirmed the validity of these results.

Using the PCR detection system adopted by Satoh et al., combined with the extraction of large plasma volumes, we were not able to detect KIs-V DNA in the blood of donors tested, suggesting an uncommon frequency in healthy persons in France. Information related to HEV status or ALT levels were not available here because both parameters are not evaluated for routine blood donor screening in France; HEV seroprevalence studies involving blood donors from northern and southwestern France indicate discrepant results (≈3%~≈52%, IgG), possibly related to serologic assay performances and/or geographic differences (4). The precise identity of KIs-V remains uncertain, but according to its extensive initial

characterization, complementary studies probably will confirm its viral origin. Molecular characterization of new full-length sequences will be needed to investigate the real genetic diversity of KIs-V and to help design optimized molecular detection systems.

The implication of KIs-V in human health remains under debate. The original publication highlighted the fact that HEV antibody-positive persons in Japan who had moderately elevated ALT levels at a prevalence of KIs-V infection that is non-negligible; such findings could suggest a link between the virus and liver dysfunctions. HEV and KIs-V also could share the same route of contamination, i.e., foods (5). Further investigations involving diverse human cohorts need to be undertaken to better understand the natural history of KIs-V in humans.

Philippe Biagini, Mhammed Touinssi, Vital Galicher, and Philippe de Micco.

Author affiliation: Etablissement Français du Sang Alpes-Méditerranée et Aix-Marseille Université, Marseille, France

DOI: http://dx.doi.org/10.3201/eid1808.120442

References

1. Satoh K, Iwata-Takakura A, Osada N, Yoshikawa A, Hoshi Y, Miyakawa K, et al. Novel DNA sequence isolated from blood donors with high transaminase levels. Hepatol Res. 2011;41:971-81.

2. Touinssi M, Brisbarre N, Picard C, Frassati C, Dussol B, Uch R, et al. Parvovirus 4 in blood donors, France. Emerg Infect Dis. 2010;16:165-6.

3. Biagini P, Bédarida S, Dussol B, Daniel L, Berland Y, de Micco P. Evolution of *Anelloviridae* strains distribution in serial blood and biopsy samples from a renal transplant patient. J Med Virol. 2012;84:96-8.

4. Mansuy JM, Bendall R, Legrand-Abravanel F, Sauné K, Miédouge M, Ellis V, et al. Hepatitis E virus antibodies in blood donors, France. Emerg Infect Dis. 2011;17:2309-12.

5. Purdy MA, Khudyakov YE. The molecular epidemiology of hepatitis E virus infection. Virus Res. 2011;161:31-9.

Address for correspondence: Philippe Biagini, UMR CNRS 7268 Equipe "Emergence et co-evolution virale," Etablissement Français du Sang Alpes-Méditerranée et Aix-Marseille Université, 27 Blvd Jean Moulin, 13005 Marseille, France; email: philippe.biagini@efs.sante.fr

Usefulness of School Absenteeism Data for Predicting Influenza Outbreaks, United States

**To the Editor:** School closure has been proposed as a strategy for slowing transmission of pandemic influenza (1). Studies of influenza A(H1N1)pdm 2009 (pH1N1) suggested that early and sustained school closure might effectively reduce communitywide influenza transmission (2,3). However, empirical evidence identifying the optimal timing of school closures to effectively reduce disease transmission after an outbreak occurs is limited.

That school absenteeism data improve school-based disease surveillance and response has been suggested (4-6). In 2009, Sasaki et al. demonstrated that the pattern of influenza-associated school absenteeism in the days before an influenza outbreak predicted the outbreak course with high sensitivity and specificity (7). However, that study used absenteeism data from Japan, which are generally not applicable to the United States, because most US absenteeism data