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Opportunities and Drawbacks of Nonstandard Body Fluid Analysis

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Opportunities and Drawbacks of Nonstandard Body Fluid Analysis Clin Chem Lab Med 2017; 55(7): 907–909 Editorial Giuseppe Lippi and Mario Plebani Opportunities and drawbacks of nonstandard body fluid analysis DOI 10.1515/cclm-2016-0862 immunochemistry assays in cancer (and other diseases) diagnostics, many drawbacks are still challenging routine The routine workout of most clinical laboratories entails, and widespread implementation (Figure 1). The collection at least for the largest part, the analysis of conventional and management of the biological samples prior to testing biological matrices such as whole blood, serum, plasma, is indeed a first and unavoidable issue. In many areas of urine and feces [1]. Nevertheless, driven by new biologi- laboratory diagnostics, preanalytical activities are highly cal discoveries and recent analytical breakthroughs (e.g. vulnerable to errors and uncertainties [3]. Nonstandard low-volume aspiration, automatic assessment of sample body fluid analysis makes no exception to this rule. The quality), the analysis of nonstandard body fluids includ- main problem here is the choice of the appropriate sample ing saliva, semen, cerebrospinal (CSF), peritoneal (i.e. type, depending on the container in which the fluid is col- ascites), pleuric, pericardic, pancreatic, synovial, cyst, lected. Although the cytological analysis of nonstandard wound, drain and washout fluids is becoming very body fluids is usually performed in blood tubes containing popular for the diagnosis and management of a kaleido- ethylenediaminetetraacetic acid (EDTA; i.e. the conven- scope of human diseases. This trend is generating new tional anticoagulant used for enumeration and classifica- and intriguing opportunities for the whole field of labora- tion of corpuscular elements), this additive is not normally tory diagnostics, but also poses important challenges for validated for standard clinical chemistry (and, occasion- reliable management of this part of non-traditional clini- ally, for immunochemistry) testing. Therefore, despite cal chemistry and immunochemistry testing. lithium-heparin or standard serum tubes are widespread The most intriguing and promising application of options for non-hematological testing in nonstandard nonstandard body fluid testing is symbolized by the body fluids, the often poor comparability of data between analysis of cancer biomarkers in fluids other than serum, these two samples types, as well as among specimens col- plasma or urine. The number of clinical studies that lected in different brands of tubes, should be regarded as have assessed many different cancer biomarkers in non- an important source of variability. The stability of many standard body fluids has exponentially grown in the past laboratory analytes in both serum and plasma has been decades, coupled by an increasing volume of routine the object of many investigations in the past 40 years [4], requests for this type of analyses placed to clinical labo- leading to the generation of reliable recommendations ratories. In an interesting article published in this issue about the most appropriate procedures for sample man- of the journal, Trimboli et al. critically review the current agement [5]. Nevertheless, little is known as yet about the scientific literature about the measurement of fine-needle most appropriate conditions of time and temperature for aspiration (FNA) endocrine markers (e.g. thyroglobulin, delayed analysis of proteins and other (cancer) biomark- calcitonin and parathyroid hormone) [2], concluding that ers in nonstandard body fluids. Measurand stability is this approach may be particularly suited for efficient and an additional concern. Many biological fluids contain relatively cheap diagnosis of thyroid/parathyroid tumors, enzymes and other proteins which may degrade or bind provided that some essential criteria are fulfilled, which to the analyte, thus artificially enhancing or decreasing its namely encompass setting adequate and standardized immunoreactivity using standard laboratory techniques preanalytical procedures, providing accurate interpreta- and ultimately producing test results which do not mirror tion criteria and appropriately addressing those cases the real concentration of the parameter in the fluid. Of par- characterized by controversial data. These important con- ticular concern is viscosity, wherein the presence of hyalu- clusions lead the way to additional thoughts on this still ronic acid and other substances in nonstandard body fluids largely debated issue. may cause aspiration errors or interference throughout the Beside the increasing clinical significance of non- analytical process. Pre-treatment with hyaluronidase may standard body fluid analysis using clinical chemistry and be effective to overcome most of these problems [6], but yet 908 Lippi and Plebani: Opportunities and drawbacks of nonstandard body fluid analysis Nonstandard body fluids: and before clinical practice implementation. This is also - Saliva - Semen an essential requirement for laboratory accreditation - Cerebrospinal according to the International Standard ISO 15189 [10]. - Peritoneal (i.e. ascites) - Pleuric Quality control testing is an essential part of laboratory - Pericardic diagnostics, entailing both the use of internal quality - Pancreatic - Synovial controls and participation to external quality assessment - Cyst - Wound Drawbacks (EQA) schemes [11]. Unlike conventional serum or plasma - Drain - Sample type analysis, analytical performance specifications and fluid- - Washout - Analyte and sample stability in nonstandard body fluids - … matched quality control materials are unavailable for - Validation of analytical techniques in nonstandard biological matrices nonconventional laboratory testing. - Lack of nonstandard fluid-matched quality control materials The lack of reliable reference ranges of many analytes - Viscosity in nonstandard biological fluids is probably the most criti- - Definition of reference ranges in health and disease cal issue. Many reasonable aspects lead us (and others) - Expert interpretation of data to believe that this target is virtually unreachable [12]. Figure 1: Leading unresolved issues in nonstandard fluid analysis. The main factors impairing the reliability of interpret- ing results of various analytes in nonstandard biologi- cal fluids include the heterogeneity of fluid production in many disease conditions (i.e. ascites, cysts, wounds poses some challenges about assay reliability in specimens and synovial infections among others), the stage of the treated with this addictive. The possible centrifugation of disease, the procedure used for collecting specimens, the the specimen is another unresolved issue (e.g. centrifuga- dilution factor and the type of the washout buffer. As it is tion force, time, temperature, etc.). Importantly, additional rather unlikely that a reference range will ever be associ- preanalytical factors were found to strongly impact the ated with results of nonstandard fluid analysis in the lab- quality of fecal immunochemical tests, thus reinforcing oratory report, interpretive information and counseling the advice of selecting appropriate collection devices and should always accompany the data. adopting specific protocols for sample collection and han- The Clinical and Laboratory Standards Institute dling [7, 8]. (CLSI) document C49-A, devoted to provide expert guid- The vast majority of clinical chemistry tests and ance about the analysis of body fluids [13], raises three immunoassays have been validated for serum and/or crucial limitations, i.e. (i) the lack of validation of com- plasma analysis by the different manufacturers. Off-label mercial assays for analysis of fluids other than whole body fluids is a concept generally referred to any type of blood, serum or plasma; (ii) the need to dedicate specific biological fluid that is different from those reported by laboratory resources for this type of testing; and (iii) the manufacturers of a Food and Drug Administration (FDA)- absence of reliable interpretative criteria (essentially, the cleared assay within the so-called “Intended Use” part lack of reliable reference ranges). These issues inevitably of the package insert [9]. Similar considerations can be call for a major involvement of laboratory professionals made for the so-called European Community-In Vitro in this type of testing, by means of validating commer- Diagnostics (CE-IVD) marking. Little evidence has been cial assays for the current “off-label” use and providing provided so far that results of many immunoassays are appropriate interpretive information about test results. reliable using sample matrices largely differing from Despite nonstandard body fluid analysis should be serum or plasma in terms of pH, proteins, ions, lipids and regarded as a promising and intriguing perspective, espe- potential interfering substances, wherein the antigen- cially in cancer diagnostics, the many current drawbacks antibody binding conditions have been optimized for necessitate supranational regulatory efforts aimed to set use under certain operating conditions, which are not a number of reliable criteria to help gathering the most always reproducible using different matrices. Moreover, safe and clinically usable information from this type on the conventional analytical performance obtained in nonconventional testing. serum or plasma (i.e. especially referring to limit of detec- tion, analytical sensitivity, linearity, imprecision) cannot Author contributions: All the authors have accepted be systematically transferred
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