Dried-Blood Spots: A Cost-Effective Field Method for the Detection of Chikungunya Virus Circulation in Remote Areas

Background In 2005, there were outbreaks of febrile polyarthritis due to Chikungunya virus (CHIKV) in the Comoros Islands. CHIKV then spread to other islands in the Indian Ocean: La Réunion, Mauritius, Seychelles and Madagascar. These outbreaks revealed the lack of surveillance and preparedness of Madagascar and other countries. Thus, it was decided in 2007 to establish a syndrome-based surveillance network to monitor dengue-like illness. Objective This study aims to evaluate the use of capillary blood samples blotted on filter papers for molecular diagnosis of CHIKV infection. Venous blood samples can be difficult to obtain and the shipment of serum in appropriate temperature conditions is too costly for most developing countries. Methodology and principal findings Venous blood and dried-blood blotted on filter paper (DBFP) were collected during the last CHIKV outbreak in Madagascar (2010) and as part of our routine surveillance of dengue-like illness. All samples were tested by real-time RT-PCR and results with serum and DBFP samples were compared for each patient. The sensitivity and specificity of tests performed with DBFP, relative to those with venous samples (defined as 100%) were 93.1% (95% CI:[84.7–97.7]) and 94.4% (95% CI:[88.3–97.7]), respectively. The Kappa coefficient 0.87 (95% CI:[0.80–0.94]) was excellent. Conclusion This study shows that DBFP specimens can be used as a cost-effective alternative sampling method for the surveillance and monitoring of CHIKV circulation and emergence in developing countries, and probably also for other arboviruses. The loss of sensitivity is insignificant and involved a very small number of patients, all with low viral loads. Whether viruses can be isolated from dried blood spots remains to be determined.

[1]  Alexander Kerner,et al.  Detection of Puumala and Rift Valley Fever virus by quantitative RT-PCR and virus viability tests in samples of blood dried and stored on filter paper. , 2011, Journal of virological methods.

[2]  Y. Leo,et al.  Chikungunya Outbreak, Singapore, 2008 , 2009, Emerging infectious diseases.

[3]  O. Marais Un impact sanitaire similaire de l’épidémie de chikungunya à la Réunion et à Mayotte , 2009 .

[4]  J. Meynard,et al.  Dengue-3 outbreak in Paraguay: investigations using capillary blood samples on filter paper. , 2008, The American journal of tropical medicine and hygiene.

[5]  M. Ratsitorahina,et al.  Outbreak of Dengue and Chikungunya Fevers, Toamasina, Madagascar, 2006 , 2008, Emerging infectious diseases.

[6]  F. Pagés,et al.  Circulation of Chikungunya virus in Gabon, 2006–2007 , 2008, Journal of medical virology.

[7]  P. Shearer,et al.  Elimination of False-Positive Polymerase Chain Reaction Results Resulting from Hole Punch Carryover Contamination , 2008, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[8]  K. Le Roux,et al.  Molecular and serological diagnosis of Chikungunya virus infection. , 2007, Pathologie-biologie.

[9]  R. Romi,et al.  Infection with chikungunya virus in Italy: an outbreak in a temperate region , 2007, The Lancet.

[10]  R. Romi,et al.  An outbreak of chikungunya fever in the province of Ravenna, Italy. , 2007, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[11]  F. Nazé,et al.  Development of a sensitive real-time reverse transcriptase PCR assay with an internal control to detect and quantify chikungunya virus. , 2007, Clinical chemistry.

[12]  J. Meynard,et al.  Use of Capillary Blood Samples as a New Approach for Diagnosis of Dengue Virus Infection , 2007, Journal of Clinical Microbiology.

[13]  S. Seneviratne,et al.  Fever epidemic moves into Sri Lanka , 2006, BMJ : British Medical Journal.

[14]  S. Higgs The 2005-2006 Chikungunya epidemic in the Indian Ocean. , 2006, Vector borne and zoonotic diseases.

[15]  V. Ravi,et al.  Re-emergence of chikungunya virus in India. , 2006, Indian journal of medical microbiology.

[16]  M. Guzmán,et al.  PCR detection of dengue virus using dried whole blood spotted on filter paper. , 2005, Journal of virological methods.

[17]  H. Guzmán,et al.  Duration of infectivity and RNA of Venezuelan equine encephalitis, West Nile, and yellow fever viruses dried on filter paper and maintained at room temperature. , 2005, The American journal of tropical medicine and hygiene.

[18]  C. Peyrefitte,et al.  Epidemic resurgence of Chikungunya virus in democratic Republic of the Congo: Identification of a new central African strain , 2004, Journal of medical virology.

[19]  F. Lahulla,et al.  Use of whole blood dried on filter paper for detection and genotyping of measles virus. , 2004, Journal of virological methods.

[20]  L. T. Figueiredo,et al.  Usefulness of blood and urine samples collected on filter paper in detecting cytomegalovirus by the polymerase chain reaction technique. , 2001, Journal of virological methods.

[21]  A. Osterhaus,et al.  Combination of Reverse Transcriptase PCR Analysis and Immunoglobulin M Detection on Filter Paper Blood Samples Allows Diagnostic and Epidemiological Studies of Measles , 2001, Journal of Clinical Microbiology.

[22]  M. Greiner,et al.  Principles and practical application of the receiver-operating characteristic analysis for diagnostic tests. , 2000, Preventive veterinary medicine.

[23]  J. Cox-Singh,et al.  Increased sensitivity of malaria detection by nested polymerase chain reaction using simple sampling and DNA extraction. , 1997, International journal for parasitology.

[24]  L. Fiore,et al.  Comparison of capillary blood versus venous blood samples in the assessment of immunity to measles. , 1996, Journal of virological methods.

[25]  J. Brody,et al.  Use of blood collected on filter paper disks in neutralization tests for poliovirus antibody. , 1965, Public health reports.

[26]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[27]  J. Héraud,et al.  Sentinel surveillance system for early outbreak detection in Madagascar , 2010, BMC public health.

[28]  A. Nordgren,et al.  Quantitation of RNA decay in dried blood spots during 20 years of storage , 2009, Clinical chemistry and laboratory medicine.

[29]  Outbreak and spread of chikungunya. , 2007, Releve epidemiologique hebdomadaire.

[30]  M. Al-Ahdal,et al.  Enzyme-linked immunosorbent assay for the detection of anti-Giardia specific immunoglobulin G in filter paper blood samples. , 1993, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[31]  F. Deinhardt,et al.  Markers of hepatitis viruses A and B: direct comparison between whole serum and blood spotted on filter-paper. , 1985, Bulletin of the World Health Organization.

[32]  H. Mirchamsy,et al.  The use of dried whole blood absorbed on filter-paper for the evaluation of diphtheria and tetanus antitoxins in mass surveys. , 1968, Bulletin of the World Health Organization.

[33]  M. P. Cunningham,et al.  The indirect fluorescent antibody technique applied to dried blood, for use as a screening test in the diagnosis of human trypanosomiasis in Africa. , 1967, Transactions of the Royal Society of Tropical Medicine and Hygiene.