Comparative sensitivity and inhibitor tolerance of GlobalFiler® PCR Amplification and Investigator® 24plex QS kits for challenging samples

Elwick, Kyleen; Mayes, Carrie; Hughes‐Stamm, Sheree

doi:10.1016/j.legalmed.2018.01.006

Cited by 23 publications

(12 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…5,17 GF-IQC with half-volume reactions produced complete profiles with 0.125 ng of input DNA and there was no difference in sensitivity from that with full volume reactions. 5,13,18 P-6C with halfvolume reactions also produced complete profiles with 0.125 ng input DNA. This was consistent with full volume reactions reported by Cisana et al 19 but differed from the report by Feng et al 20 the groups reported complete profiles with DNA amounts of 0.125 and up to 62.5 ng, respectively.…”

Section: Discussionmentioning

confidence: 97%

“…Melanin, hematin and humic acid were dissolved in an appropriate solvent (melanin: 0.5 M NH 4 OH; hematin: 0.1 M NaOH; humic acid: nuclease-free water). 13 Each inhibitor, along with an optimal amount of input control DNA 1224, was used as half-volume for amplification. Then, the samples were tested with an ABI 3500 Genetic Analyzer and examined at 175 RFU.…”

Section: Inhibitor Studymentioning

confidence: 99%

See 1 more Smart Citation

Comparison of the Efficiency of GlobalfilerTM IQC PCR Amplification Kit and Powerplex® Fusion 6C System with Half-Volume Reactions for Forensic Trace DNA Testing

Nattawoot Saokaew,

Kewalee Junpan,

Wawkan Duangshatome

et al. 2022

Indian Journal of Forensic Medicine & Toxicology

View full text Add to dashboard Cite

Two major challenges in daily and routinely forensic genetics test are samples with low or very low quantities of DNA and high cost of processing. To overcome these obstacles, a reduction in reaction volume to half would offer a substantial benefit, but even so it should provide complete genetic profiles without conceding the quality of the results. The aim of this study is to test the robustness of two commercially available kits, GlobalFiler™ IQC PCR Amplification Kit (GF-IQC) and PowerPlex ® Fusion 6C System (P-6C), with half volume reaction in accordance with SGWDAM guidelines. GF-IQC showed minimum and stochastic threshold of 45 and 610 RFU, while P-6C demonstrated 60 and 670 RFU of those values respectively. The P-6C can tolerate to many inhibitors including melanin, hematin, and humic acid. Both kits showed the same performance on case work profiling and the sensitivity of allele detection of P-6C was not significantly different from that of . The results of this study demonstrated that GF-IQC and P-6C with half volume reaction can produce good quality profiles and could be applied for forensic DNA examination.

show abstract

Section: Discussionmentioning

confidence: 97%

Section: Inhibitor Studymentioning

confidence: 99%

Comparison of the Efficiency of GlobalfilerTM IQC PCR Amplification Kit and Powerplex® Fusion 6C System with Half-Volume Reactions for Forensic Trace DNA Testing

Nattawoot Saokaew,

Kewalee Junpan,

Wawkan Duangshatome

et al. 2022

Indian Journal of Forensic Medicine & Toxicology

View full text Add to dashboard Cite

show abstract

“…In addition, biological samples such as blood, urine and hair contain various PCR inhibitors including hematin, urea and melanin that may be co-extracted with the DNA [47][48][49]. These PCR inhibitors can negatively affect the DNA typing results by binding to the polymerase or interacting with DNA, which results in partial or complete inhibition of PCR [50][51][52][53][54].…”

Section: Stability Studymentioning

confidence: 99%

Developmental Validation of the Novel Six-Dye GoldeneyeTM DNA ID System 35InDel Kit for Forensic Application

Yang

et al. 2021

Forensic Sciences Research

View full text Add to dashboard Cite

Insertion/deletion polymorphisms (InDels) have been treated as a prospective and helpful genetic marker in the fields of forensic human identification, anthropology and population genetics for the past few years. In this study, we developed a six-dye multiplex typing system consisting of 34 autosomal InDels and Amelogenin for forensic application. The contained InDels were specifically selected for Chinese population with the MAF ≥ 0.25 in East Asia, which do not overlap with the markers of Investigator ® DIPplex kit. The typing system was named as Goldeneye TM DNA ID System 35InDel Kit, and a series of developmental validation studies including repeatability/reproducibility, concordance, accuracy, sensitivity, stability, species specificity and population genetics were conducted on this kit. We confirmed that the 35InDel kit is precise, sensitive, species specific and robust for forensic practice. Moreover, the 35InDel kit is capable of typing DNA extracted from forensic routine case-type samples as well as degraded samples and mixture samples. All markers are proved to be highly polymorphic with an average observed heterozygosity (He) of 0.4582. The combined power of discrimination (CPD) is 0.999 999 999 999 978 and the combined power of exclusion in duos (CPE D ) and trios (CPE T ) are 0.978 837 and 0.999573, respectively, which are higher than those of the Investigator® DIPplex kit. Thus, the Goldeneye TM DNA ID System 35InDel kit is suitable for forensic human identification and could serve as a supplementary typing system for paternity testing.

show abstract

“…A major concern with removing the extraction step is that inhibitors of PCR remain in the sample. This may be circumvented by the use of inhibitor-resistant STR kits, such as the Promega PowerPlex 21 PCR Amplification System, and Thermo Fisher's GlobalFiler PCR Amplification Kit (Brito et al, 2015;Elwick, Mayes, & Hughes-Stamm, 2018;Gray, Crowle, & Scott, 2014). In addition to autosomal STR profiles, complete Y-STR profiles have also been produced from blood and saliva samples deposited on cigarette butts, drinking straws, grass, leaves, woodchips, and fabrics without extraction and quantification (Dargay & Roy, 2016).…”

Section: Dna Profilingmentioning

confidence: 99%

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Lynch

Fleming

2019

WIREs Forensic Science

View full text Add to dashboard Cite

Short tandem repeat (STR) profiling is frequently carried out on biological stains in forensic casework, and the application of messenger RNA (mRNA) body fluid identification is becoming more prevalent. Increasing constraints on analysis time, resources, an increase in the number of samples for analysis, and a large backlog of cases creates significant demand for quicker methods that direct polymerase chain reaction (PCR) can help overcome. Direct PCR bypasses DNA and/or RNA extraction, and in some cases, quantification by adding the sample or substrate punch to the amplification reaction. This method is currently used in some laboratories on reference samples. Direct PCR is also a component of portable devices, which could improve efficiency even further by producing results at the scene, and enabling triaging of samples for further testing. Portable devices are currently used for STR profiling of reference samples but not for mRNA profiling. Various concerns regarding the reliability of the method and equipment (portable devices) must first be addressed before incorporation into forensic casework. This review discusses the limitations and factors that need to be considered for direct PCR to be used for the analysis of casework samples, including being used for mRNA body fluid identification. This article is categorized under: Forensic Biology > Forensic DNA Technologies Forensic Biology > Body Fluid Identification Forensic Biology > Applications of RNA

show abstract

Comparative sensitivity and inhibitor tolerance of GlobalFiler® PCR Amplification and Investigator® 24plex QS kits for challenging samples

Cited by 23 publications

References 28 publications

Comparison of the Efficiency of GlobalfilerTM IQC PCR Amplification Kit and Powerplex® Fusion 6C System with Half-Volume Reactions for Forensic Trace DNA Testing

Comparison of the Efficiency of GlobalfilerTM IQC PCR Amplification Kit and Powerplex® Fusion 6C System with Half-Volume Reactions for Forensic Trace DNA Testing

Developmental Validation of the Novel Six-Dye GoldeneyeTM DNA ID System 35InDel Kit for Forensic Application

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Contact Info

Product

Resources

About