INSTRUCTION. Guidelines for samples submission and analytical requests
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1 GUADELOUPE SITE DE DUCLOS CRVC INSTRUCTION Guidelines for samples submission and analytical requests I-TE-06 Version 07 Du 25/08/17 This guideline aims to inform the client on the overall analysis provided by the CIRAD laboratory (Duclos location) and to follow the guidance of sampling, storage and delivery samples. This document describes also the method to limit biological hazards during the freight to the destination and keep good preservation of the samples. These guidelines are addressed to DAAF, to chief veterinary officers, laboratories, veterinarians, farmers or any other client requesting a sample analysis. Rédaction : RE Nom : Signature : 21/08/2017 Vérification : Responsable CRVC Validation : RQSE Nom : Nom : Signature : Signature : 21/08/ /08/2017 Page 1 sur 6
2 ANALYTICAL REQUEST PRIOR to sending any samples, the client must inform the CIRAD laboratory by fax, phone or of his/her intention. By doing so, the client will be ensured that a qualified person (biologist responsible or responsible of assay technician) will be present at the CIRAD laboratory to receive the delivery. In the other case, the customer is taking the risk that the package will be send back if there is no qualified person. Head office : Secrétariat : Emmanuel Albina Tél : Cell : emmanuel.albina@cirad.fr Yolaine Vincent Tel : Fax : Technicians : Rosalie Aprelon Valérie Pinarello Kaïssa Plaisir CIRAD-Duclos Prise d eau, Petit Bourg Head deputy: Quality manager : Table 1 lists all the diseases and techniques used by the laboratory. Jennifer Pradel Tél : Cell : jennifer.pradel@cirad.fr Christian Sheikboudou Theses analyses are covered by the accreditation N (available on website on All right reserved by the COFRAC brand. The accreditation reference is allowed if it does not compromise the accreditation of the laboratory and the COFRAC. Table 1: Laboratory Analyzes and Pricing Price (without taxes) Disease Pathogen Specie Technique Sample Cowdriosis/ (M gene) (H5 gene) (H7 gene) Newcastle disease Pandemic influenza ph1n1 Babesiosis Ehrlichiosis Ehrlichia ruminantium Virus Avian influenza virus Newcastle virus Pandemic swine H1N1 influenza virus Anaplasma marginale Anaplasma ovis Anaplasma phagocytophilum Anaplasma platys Babesia bovis Babesia bigemina Bovine, Caprine, Ovine Equines, Bird Whole blood, tick, tissue RT- Whole blood, tissue Real time RT- Serum, Whole blood, Cerebrospinal fluid, tissue Bird Real time Tissue, swab Bird Swine Bovine Caprine, Ovine Real time RT- Real time RT (M gene) Real time RT- (N1 gene) Tissue, swab Tissue, swab Whole blood Whole blood Equine Whole blood Dog Bovine Whole blood Real time Whole blood Real time Whole blood Ehrlichia canis Dog Whole blood Ehrlichia sp Bovine, Caprine, Ovine, Equine Whole blood Page 2 sur 6
3 ANALYTICAL REQUEST FORM The client will have to fill and include the analytical request form E-TE-02 Follow-up of Samples in the samples package to be analyzed or at the latest 24 hours after the delivery otherwise it will delay all the analysis process. The sample submission will be officially validated only after reception of the sample follow-up form and register in numbering system of the laboratory. The biological samples could be inactivated (by chemical or temperature process) or potentially infected. In the first condition, the sender laboratory must provide a certificate including the inactivation methods signed by the laboratory manager who has done the inactivation. In the second condition, the samples will be considered as infectious materials and will be handled following IATA guidelines. By air TRANSPORT OF SAMPLES The client should include an Aircraft Freight Letter with his name, phone number and address, those from the consignee laboratory and the samples description and characteristics (UN or non-dangerous samples) and their value. BEFORE any research or diagnosis animal samples shipment from non-european Union countries, CIRAD (destination laboratory) must obtain an importation permit from to French veterinary services which need a complete description of the samples (number, nature, origin). CIRAD will transmit this import permit to the customer who should include it in the shipment. This request is related to regulation policies published in the Official Journal of the French Republic and is available upon request (EXT 102 EB / TD-TE). IATA regulations Inactivated samples shipping do not need specifics conditions related to IATA regulation. The IATA rules must be followed to freight any infectious substances by plane. Infectious substances are listed in the 6.2 section of the IATA rules and there are two different categories: - Category A= If there is any problem with the container (failure, rupture, break), the infectious substances could affect humans or animals. Human pathogens are classified as UN2814 and Animals pathogens UN Category B= Infectious substances which are not classified the category above are classified as UN3373. Depends on the classification of the infectious samples, the freight and the packaging will varied. The flow chart below summarizes the guidelines. More details are described in the P-TE-04 document (available upon request). UN2814 Human Pathogen Category A Contagious pathogen INFECTIOUS SUBSTANCES UN2900 Animal Pathogen Category B Non contagious pathogen UN3373 Biological substances Recommendations: Send samples at the beginning of the week & avoid holiday period Provide tracking number as soon as possible: FEDEX or other courier company Triple packaging: sealed primary container, sealed second container with absorbent and external tertiary packaging Limited quantity required Page 3 sur 6
4 Courier Company Transportation is usually done by the company: - FEDEX Company is the main company able to transport diagnosis specimens in the Caribbean except Cuba; - DHL in Surinam - AMERIJET for the Guyana. Each laboratory has to contact his local agency to make sure that they can take in charge 6.2 section biological hazards, diagnostic samples. Depend on the company the conditions of shipment authorization could vary. For inactivated or potentially infectious substances to be freight obligatory documents must be in the package. See above guidelines check list concerning the shipment and analysis: GUIDELINES Prior contact the laboratory (fax, or by phone) Fill in the E-TE-02 form and include in the package or send at latest 24 h after reception of the shipment Sample description form (specie, age, sex ) Import permit for samples for non-european countries (provided by CIRAD) Inactivated samples Certificate (established by the sender laboratory) Air Craft Freight Letter with name and coordinates of the sender and recipient and description of samples Follow the samples preservatives guidelines from 4 & 5 chapters of this document Individually identification of samples Infectious Substance Declaration UN 2814, 2900 ou 3373 classifications Triple containers packaging PRICING See Table 1. Rates are quoted excluding taxes: a tax of 8.50% applies. For requirements involving more than 200 samples, a prior agreement will be requested and appropriate tariffs may be considered. Recipient RESULTS RELEASE The client is the person who requests the analysis (E-TE-02 form) and he will be the unique recipient of the results and will be invoiced. If the customer asks to another laboratory to pack and send the samples, the result will be uniquely sent to him except if there is an agreement between the client and the laboratory which did the shipment. For any disease classified in sanitary danger category 1 or 2 and submitted to control measures in the DFA, the client is the DAAF of the concerned department. It concerns for example the avian influenza and heartwater for Martinique. In this case, results will not be sent to the person who did the sampling or the veterinarian that would possibly have done the initial analysis request, except if DAAF did gave an prior agreement for results dissemination. In the contrary case, the person who requested the test is invited to contact DAAF. Deadlines Results will be released to the client by or fax with the E-TE-09. The time for results release will depend on the ongoing research activities in the lab and the personnel available. A deadline can be provided upon request at the time of the analysis request. Generally, the expected timeframes are shown in Table 2. Table 2: Reporting Delays Page 4 sur 6
5 Real time temps for avian influenza: Real time temps Newcstle: : ELISA : : ELISA : 48 hours for a partial diagnosis based on M gene in case of clinical suspicion 48 hours 30 days. 7 days in case of suspicion in free heartwater area 30 days 7 days 30 days, for epidemiological study 7 days for a clinical suspicion 15 days for animal importation. Analytical request for Avian influenza diagnosis is the CIRAD laboratory priority, samples will be process within 24h after reception, in case of emergency (weekend and holiday period) there will be a technical permanence to do analysis. If exceptionally, the CIRAD laboratory cannot process the samples (due to limited amount of material or staff restriction), the sample submission and analytical request will be transferred to an adequate reference laboratory. In this case, the CIRAD laboratory informs the client (by mail) and requests his/her authorization (by mail). CIRAD can provide advices and interpretations on the result report in order to facilitate the understanding of the sample analysis by the client. CIRAD may also produce advices and interpretations out of the context of test reports. Partial release of the results When several analysis are requested if one of them is urgent (clinical suspicion of Avian Influenza for example) the laboratory can provide a partial result. When the final report is released, the customer must destroy or return to the CIRAD laboratory the initial report that was the subject to the amendment. The CIRAD laboratory is authorized to use non identified results obtained from the analysis for technical and scientific purposes (mainly for scientific publication). Unsatisfied customer (refusal of analysis or results delay for examples) can send a complaint to CIRAD laboratory either by phone or by written statement. CHARACTERISTICS OF SAMPLES AND STORAGE The table 3 below shows the guidelines for sampling, storage and shipment depending on the samples. If the storage and shipping recommendations for samples are not respected, the CIRAD laboratory can refuse to perform the analysis; still, if the analysis is done, it will be performed under derogation. Samples for the serology analysis can be: - blood which must be on a dry tube and will be centrifuge - serum. Samples for analysis can be: - Whole blood, - Organ - vectors (ticks, mosquitos). The sample quality must be good to process the analysis (container integrity, without contamination, stain, physic-chemical quality: smell, aspect, color, etc ). In a doubt, a discussion between RE and RTB will be organized. Any suspicion of bacterial or fungus contamination (green or white color, bacterial or fungi, colony or strong smell ) the samples will be not accepted. In a particular case of swabs, the quality of the sample can t be jugde. The quantity of the sample must be enough. Refusal has to be considered: - all the conditions presented in the table below are not respected: nature of the sample, quantity, preservative and time limit. - the samples display unusual physico-chemical qualities (smell, aspect, color) In any case, the tubes or the swabs containing the samples must be clearly and individually identified. Table 3: Guidelines for storage and preservation of samples 1 Dilution in ethanol: 3 volumes of blood for 1 volume of ethanol at ethanol, RNA later and antibiotic medium are provided by the laboratory. Page 5 sur 6
6 Samples Method Disease Characteristic of samples Blood Serum Serology 1 Serology Cowdriosis Ehrlichiosis Babesiosis SAMPLE PRESERVATION CONDITIONS 1mL minimum On EDTA or heparinate tubes 1mL minimum On dry tube 500 µl minimum of serum obtained from coagulated blood and centrifuged 10 min between 400 and 800 g Storage conditions prior to lab delivery Maximum time between sample collection and delivery Shipment condition +4 C 4h +4 C (with freeze packs) -20 C 15 days +4 C (with freeze packs) Ambiant Temperature in ethanol days Ambiant Temperature +4 C in ethanol 12 3 months Ambiant Temperature -20 C in ethanol 12 6 month Ambiant Temperature +4 C 4 h +4 C (with freeze packs) + 4 C (can stay 2 hours max at ambiant temperature) 24h +4 C (with freeze packs) - 20 C 3 months +4 C (with freeze packs) Plasma Blood collected on EDTA tube Cephalo-spinal liquid Organ minimum 500 µl (brain) (Liver, heart, kidney, spleen, lung ) Newcastle (brain) (Liver, heart, kidney, spleen, lung) Ticks Swabs Swine Influenza Newcastle disease minimum 100 mm 3 minimum 100 mm 3 Nymph or alive adult tick Nymph or dead adult tick Tracheal or cloacal swabs (Avian) and nasal swabs (swine) on sick or dead animals Tissue and organ on dead animals + 4 C 5 days +4 C (with freeze packs) - 80 C 3 months <-20 C (dry ice of freeze packs -80 C) +4 C <24h +4 C (with freeze packs) -80 C >24h <-20 C (dry ice of freeze packs -80 C) -20 C 48h <-20 C (dry ice of freeze packs -80 C) +4 C in RNA later 2 24h +4 C (with freeze packs) -20 C 48 h +4 C (with freeze packs) Ambiant Temperature in ethanol days Ambiant Temperature +4 C in ethanol 12 3 months Ambiant Temperature -20 C in ethanol 12 6 months Ambiant Temperature Ambiant temperature use of plastic tube with aeration Ambiant temperature and storage in ethanol 12 3 days Ambiant Temperature 3 months Ambiant Temperature +4 C in antibiotic media 2 24 h +4 C (with freeze packs) -20 C in antibiotic media 2 72 h <-20 C (dry ice of freeze packs -80 C) Page 6 sur 6
7 Page 7 sur 6
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