Immunochromatographic detection of human hemoglobin from deteriorated bloodstains due to methamphetamine contamination, aging, and heating

Murahashi, Masataka; Makinodan, Mitsuru; Yui, Megumi; Hibi, Takao; Kobayashi, Motohiro

doi:10.1007/s00216-020-02802-6

Cited by 5 publications

(9 citation statements)

References 14 publications

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“…As already stated, also the aging of the stains is a potential source of false-negative results because environmental factors can promote the modification/denaturation of the antigenic properties of the targets or make the targets insoluble and therefore un-accessible to the assay [17,24,26,35,45,47]. In line with this, a lower sensitivity leading to false negative results was observed in the analysis of year-aged bloodstains stored at room temperature [26].…”

Section: Ageing Of the Samplementioning

confidence: 73%

“…It has to be noted, however, that the methodology used to establish the sensitivity (that is, the limit of detection, LOD) is not fully described because scarce/null details are provided of the blood samples used in those internal validation procedures. In addition, the LOD value is not provided at all in some kits developed for clinical purposes only (this applies, for example, to the OC-Hemocatch tests, which were developed to detect occult blood in feces but are widely employed in East Asia even for forensic purposes [45][46][47]. Certified hHb was used only in the SERATEC ® HemDirect kit [33], whereas other kits seem to have used fresh blood from unknown donors.…”

Section: Sensitivity Of the Ic Assaysmentioning

confidence: 99%

“…In both circumstances, the amount (i.e., the volume) of blood needed for a positive outcome could be higher than that required from fresh blood samples because a complete extraction of the antigenic target from a stain cannot be assured. The hHB/Gly-A extraction could be inhibited by several factors such as heat, which can modify the solubility of the antigenic target [47]; in other cases, the intrinsic properties of the swab used for sampling the stain can interfere with the accuracy of the assay [28]. Validation studies performed on different sets of bloodstains usually evaluate and report on the aging of the samples, the storage conditions, and, sometimes, the size (in square millimeters) of the sample used for IC testing, whereas the outcome is indicated as "positive" (with +++, ++ and + arbitrary scores) in only a few cases or "negative"; as a result, it is impossible to establish any LOD for the selected IC assay.…”

Section: Sensitivity Of the Ic Assaysmentioning

confidence: 99%

“…Validation studies performed on different sets of bloodstains usually evaluate and report on the aging of the samples, the storage conditions, and, sometimes, the size (in square millimeters) of the sample used for IC testing, whereas the outcome is indicated as "positive" (with +++, ++ and + arbitrary scores) in only a few cases or "negative"; as a result, it is impossible to establish any LOD for the selected IC assay. Of great importance in forensics, bloodstains are often aged and/or exposed to environmental factors that can potentially modify/degrade the antigenic properties of the proteins [24,47]. In such cases, much higher LODs are expected, and, in some instances, false-negative results can be found (see Section 5).…”

Section: Sensitivity Of the Ic Assaysmentioning

confidence: 99%

“…In some cases, although scarce information is sometimes given on the stain size used for the assays, the evidence suggests that the degradation/modification of the antigenic epitopes could prevent them from being recognized by the antibodies for which they are targets [24]. In other cases, such as heating, the reduced solubility of the antigenic targets seems to be involved in the negative outcome of the assay [47].…”

Section: Ageing Of the Samplementioning

confidence: 99%

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Immunochromatographic Detection of Human Blood: A Forensic Review

Cuttaia,

Di Stefano,

Sorçaburu Ciglieri

et al. 2024

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Body fluid identification is fundamental in forensic science as it links a specific biological source to a genetic profile, thus providing critical clues for crime scene reconstruction. Blood is one of the most common body fluids found on the crime scene, and several strategies have been developed for its identification in recent decades. Usually, after a preliminary (or presumptive) test to determine the presence of blood (both human and non-human), a confirmatory test is needed to prove that the sample is human blood. Out of the confirmatory tests, immunochromatographic (IC) assays are the most commonly and widely used. This work gives a review of the use of commercial kits specifically developed to detect human hemoglobin or glycophorin A (a surface protein of human red cells) in forensics. Claimed sensitivity varies broadly (ranging from 0.06 to 75 nanoliters of fresh blood), but different values (as low as 0.002 nL) were found during validation procedures. Specificities are high, and the possibility of cross-reaction (with the risk of false-positive results) is so low that it can be considered negligible. False-negative results, however, can be found due to the so-called “hook effect” as well as to the target degradation/modification, which interferes with the Ag-Ab binding. In addition, the chemical compositions of the presumptive test, detergents, and washing can also promote false negative outcomes in peculiar situations. Although IC assays are rapid, inexpensive, specific, and easy to use even on the crime scene, their major limitation is represented by the destructive approach required by this kind of confirmatory test. Since the final goal of the forensic investigation is the genetic typing of a bloodstain, we will describe the strategies developed for IC assays of faint stains as well as the strategies adopted to ensure that exactly the same sample undergoes human blood identification and DNA typing.

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