False‐Positive Results with Amylase Testing of Citrus Fruits

Ricci, U.; Carboni, Ilaria; Torricelli, Francesca

doi:10.1111/1556-4029.12457

Cited by 10 publications

(8 citation statements)

References 14 publications

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“…The results of the current study are in consistent with (Old et al, 2010 andRicci et al, 2014).who stated that in humans, two main isozymes of α-amylase exist, salivary and pancreatic; old methods for saliva identification cannot distinguish between these different α-amylase isozymes. Thus, the RSID TM -Saliva kit was developed; it utilises two antihuman-salivary amylase monoclonal antibodies to detect the presence of αamylase rather than its activity which, could be compromised in aged or degraded forensic samples.…”

Section: Discussionsupporting

confidence: 92%

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Rapid Stain Biomarker : A Novel Tool for the Forensic Detection of Saliva (Effect of Time and Surface on its Reliability )

Mahmoud

Zaki

2018

Ain Shams Journal of Forensic Medicine and Clinical Toxicology

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Saliva is one of the vital fluids secreted by the human body; it can be deposited on the human skin or other materials through biting, sucking, licking and kissing. Saliva stains encountered in forensic casework is an important source of DNA; so, it is considered an important evidence for personnel identification. The rapid Stain Identification of Human Saliva (RSID TM-Saliva) bioassay is designed to detect specifically the presence of human salivary -amylase enzyme. Test development is completed within 10 minutes and can detect as little as 1l of human saliva. The detection protocol can be completely integrated into the procedures for DNA extraction and analysis. Aim: Assess the efficacy of RSID TM-Saliva strip test for the detection of human saliva under some different variables (different surfaces & different time intervals).Evaluate the quality of DNA extracted from unpreserved saliva and the possibility of its usage as evidence in forensics. Methodology: Fifty two saliva samples were collected from four volunteers. Forty eight of them were deposited over surfaces of different nature (plastic bottles, glass cup and cigarette butts) at the same time. Four saliva samples were collected directly from the floor of the mouth of each participant by a cotton swab (positive control group). All samples were left to dry for 10 min at room temperature then extracted and analyzed according to the protocol designed for (RSID TM-Saliva). Collected samples were studied as three tested groups and as control groups. Saliva samples collected from each participant were categorized into four subgroups according to the time interval of saliva extraction [zero (on the spot), 10, 20, and 30 days].Positive samples for saliva identification were subjected to DNA extraction and typing; as three autosomal primers short tandem repeats (STRs) specific to human genomic DNA (D21S11, D18S51 and CSF1PO) were amplified by polymerase chain reaction (PCR); then subjected to separation and analysis of fragments size by agarose gel electrophoresis to determine their allelic size through comparison to the standard allelic 100 bp ladder. Results: Saliva could be identified in 100% of tested samples as well as for the positive control samples, which were subjected to successful DNA extraction followed by PCR amplification of the chosen three STRs. Results of agarose gel revealed that the sizes of these PCR products were in accordance with the 100 bp DNA ladder; validating the good quality and quantity of the extracted DNA Conclusion: the current study demonstrated that non-stored saliva deposited over different surfaces for up to 30 days presents an ideal source for DNA which may be used for forensic identification. The study also evidenced that the new RSID-saliva strip test is a fast, easy, sensitive and reliable method for saliva identification over different surfaces.

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Section: Discussionsupporting

confidence: 92%

“…However, it gave positive reactions for gorilla and rat saliva, breast milk samples and citrus fruits as lemon, orange, grape fruit etc. (Ricci et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Rapid Stain Biomarker : A Novel Tool for the Forensic Detection of Saliva (Effect of Time and Surface on its Reliability )

Mahmoud

Zaki

2018

Ain Shams Journal of Forensic Medicine and Clinical Toxicology

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“…However, the RSID™‐saliva test is more costly than the Phadebas ® amylase test and ELISA requires long processing times and is more complicated to perform than the Phadebas ® test. Moreover, the RSID™‐saliva test has been reported to include false‐positive reactions to other body fluids and citrus fruits . Because this study just focused on the amylase tests, further research focused on the other saliva markers is needed to apply various forensic samples.…”

Section: Resultsmentioning

confidence: 99%

Comparison of Catalytic and Immunological Amylase Tests for Identifying of Saliva from Degraded Samples

Ohta

Noda

Sakurada

2018

Journal of Forensic Sciences

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The stability of salivary α‐amylase is a critical factor in both catalytic and immunological method‐based forensic saliva identification. This study aimed to assess the sensitivity of catalytic and immunological tests on degraded saliva samples. Degraded saliva stains were prepared by microbial decomposition using humid soil. Salivary α‐amylase activity was catalytically detected both qualitatively and quantitatively using the Phadebas® amylase test. As immunological methods, we conducted qualitative and quantitative tests using the RSID™‐saliva test and ELISA, respectively. Salivary α‐amylase activity of degraded samples (incubated at 37°C for 12 h) was significantly lower than that of controls in the quantitative tests. All the degraded samples obtained by the humid soil produced negative results in the Phadebas® tests, but showed positive results in the RSID™‐saliva test and ELISA. These results suggest that immunological tests are effective for testing degraded saliva samples that have lost their enzymatic activity.

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“…At this level of sensitivity, it is thought that the assay is detecting very low amounts of hemoglobin in these non-target fluids; thereby generating false positive reactions in regard to the biological fluid being targeted. However, unpredictable cross-reactivity with non-target molecules having similar conformational epitopes is also possible as are non-specific binding J o u r n a l P r e -p r o o f events due to extremes of pH, the presence of organic acids or other sample-specific chemical compounds [226,227]. Environmental contaminants also have the possibility of interfering with antibody binding [29].…”

Section: J O U R N a L P R E -P R O O Fmentioning

confidence: 99%