The establishment of a multiplex fluorescent polymerase chain reaction coupled with capillary electrophoresis analysis technology enables the simultaneous detection of 16 genotypes of human papillomavirus.

Background: The detection and accurate genotyping of human papillomavirus (HPV) infection is critical for preventing and effectively treating cervical cancer.

Methods: A multiplex fluorescent polymerase chain reaction (PCR) coupled with a capillary electrophoresis method was developed for the simultaneous detection of the 16 most prevalent HPV genotypes. Twenty-five pairs of primers were ultimately selected to ensure that both E and L regions of nine HPV genotypes, as well as the E regions of seven HPV genotypes could be accurately amplified.

Design and development of novel single multiplex system incorporating 26 rapidly mutating Y-STRs; 26 RM Yplex.

This article details the development of a single multiplex system amplifying 26 rapidly mutating Y-STR markers. A sequenced allelic ladder, constructed for calling alleles of all loci, is introduced. The multiplex system shows the ability to address the limitations of Y-STRs commercial kits in differentiating closely related males.

Validation of the Microreader 28A ID System: A 6-dye multiplex amplification assay for forensic application.

The Microreader 28A ID System is a new 28-plex genotyping system with 6-dye multiplex amplification, which allows the simultaneous amplification of all 20 Combined DNA Index System (CODIS) core loci (CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, FGA, TH01, TPOX, vWA, D1S1656, D2S441, D2S1338, D10S1248, D12S391, D19S433, D22S1045), plus five extended STRs loci (D6S1043, Penta D, Penta E, DYS391, SE33), 2 Y-Indels (Rs2032678, Rs771783753), and the amelogenin loci. This system can be used for forensic analyses, such as personal identification, kinship testing, scientific research, database applications, and other aspects of human genetic identification. The validation of the Microreader 28A ID System followed the "Validation Guidelines for DNA Analysis Methods (2016)" described by the Scientific Working Group on DNA Analysis Methods and the regulations published by the China Ministry of Public Security.

Developmental validation of the Microreader™ Y Prime Plus ID System: An advanced Y-STR 38-plex system for forensic applications.

Y-STR is widely used in sexual assaults and familial searches of suspects. Here, we reported a novel 38-plex STR genotyping system designed for forensic applications. Microreader™ Y Prime Plus ID System (YPP) amplifies 38 loci in one reaction, including 29 loci from commonly used Yfiler® Plus PCR Amplification Kit & PowerPlex® Y23 System (DYS393, DYS570, DYS19, DYS392, DYS549, Y GATA H4, DYS460, DYS458, DYS481, DYS635, DYS448, DYS533, DYS449, DYS456, DYS389I, DYS390, DYS389Ⅱ, DYS438, DYS391, DYS439, DYS437, DYS385a/b, DYS643, DYS518, DYS576, DYF387S1a/b, and DYS627), 6 commonly used loci for the Y-STR database (DYS444, DYS447, DYS596, DYF404a/b, DYS527a/b, DYS557) and one Y-indel specific for the Chinese population.

Development and validation of a multiplex 19 X-chromosomal short tandem repeats typing system for forensic purposes.

X-chromosome short tandem repeat (X-STR) markers are a powerful complementary system used for paternity and forensic casework. This study presents the development and validation of a new highly efficient multiplex-fluorescent-labeled 19 X-STR typing system, including DXS10079, DXS101, DXS10135, DXS10162, DXS6795, DXS6800, DXS6803, DXS6807, DXS6809, DXS6810, DXS7133, DXS7423, DXS981, DXS9902, DXS9907, GATA165B12, GATA172D05, GATA31E08 and HPRTB along with sex-typing locus, amelogenin. The system was validated according to guidelines issued by the Scientific Working Group on DNA Analysis Methods.

Validation of the Microreader 40Y ID System: a Y-STR multiplex for casework and database samples.

Y-chromosome-specific short tandem repeat loci (Y-STRs) are commonly analysed in forensic science for paternity testing, familial searches, and, in sexual assault cases, to determine male DNA identity from mixed sources with high background female DNA content. The Microreader 40Y ID System is a six-dye multiplex amplification kit that contains 17 Y-STR loci from the Yfiler Plus PCR Amplification Kit and the powerplex Y23 system (DYS19, DYF385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS549, DYS635(Y GATA C4), DYS643, Y GATA H4, DYS460, DYS481, DYS533, DYF387S1, DYS449, DYS518, DYS570, DYS576, and DYS627), plus six high polymorphic loci (DYS444, DYS447, DYS557, DYS596, DYS527 a/b) as well as 4 additional candidate Y-STR loci (DYS593, DYF404S1, DYS645) and a Y-Indel loci (Rs2032678), thereby providing greater efficiency, compatibility, and accuracy. The Microreader 40Y ID System can directly amplify markers from blood or saliva on filter paper or FTA cards, without template extraction or purification, and can also be used for extracted DNA templates.

Validation of the Microreader™ 29Y Prime ID system for forensic use.

Currently, Y-short tandem repeat loci (Y-STRs) have been increasingly used in the forensic field, particularly in investigations of sexual assault, determination of paternity and male lineage studies because of the characteristics of male-only and paternal inheritance. The Microreader™ 29Y Prime ID system is a 29-plex Y-STR genotyping system that amplifies 17 widely used commercial loci (DYS570, DYS546, DYS460, DYS458, DYS635, DYS533, DYS448, DYS627, DYS456, DYS576, DYS449, DYS437, DYS643, DYS518, DYF387S1 a/b, and a sexual locus Y GATA H4), European recommended 7 single-copy "minimal haplotypes" (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, and DYS385a/b) and 2 additional loci (DYS438 and DYS439) recommended by The Scientific Working Group on DNA Analysis Methods (SWGDAM). The Microreader™ 29Y Prime ID system was validated according to the guidelines of "Validation Guidelines for DNA Analysis Methods (2016)" described by the Scientific Working Group on DNA Analysis Methods (SWGDAM), including PCR-based, sensitivity, precision and accuracy evaluation, stutter percentage and peak height ratio, inhibitors, species specificity and DNA mixture studies.

Developmental validation of the Microreader™ 20A ID system.

The Microreader™ 20A ID system is designed for forensic applications such as personal identification, parentage testing, and research. It includes 13 combined DNA index system (CODIS) short tandem repeat (STR) loci (CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, and D21S11), three expanded CODIS STR loci (D12S391, D19S433, and D2S1338), three non-CODIS STR loci (D6S1043, Penta D, and Penta E), and the amelogenin locus in one reaction with a six-dye fluorescent (FAM, HEX, TAMAR, ROX, PUR, and QD550) analysis system. In this study, the Microreader™ 20A ID system was validated according to the Scientific Working Group on DNA Analysis Methods validation guidelines for forensic DNA Analysis methods and Chinese national standard, including PCR-based studies, sensitivity study, precision, and accuracy evaluation, stutter calculation, inhibitor tests, species specificity, and DNA mixture studies.

Inconsistent genotyping call at DYS389 locus and implications for interpretation.

The male-specific Y chromosome short tandem repeat (STR) locus is used widely in forensic case, which are useful molecular tool to providing the biological evidence for male/female mixture and paternal lineage cases. The Y-STR analysis has been greatly facilitated by advent of commercial multiplex kit. However, even with well-designed robust multiplex kit, abnormal genotyping profile may be observed when encountering with mutations, such as deletion/duplication within the target region or mutation at the primer binding site.

Validation of the Microreader™ 23sp ID system: A new STR 23-plex system for forensic application.

Microreader™ 23sp ID system is a new 23-plex STR genotyping system that amplified 21 non-CODIS STR loci (D6S477, D18S535, D19S253, D15S659, D11S2368, D20S470, D1S1656, D22-GATA198B05, D7S3048, D8S1132, D4S2366, D21S1270, D13S325, D9S925, D3S3045, D14S608, D10S1435, D12S391, D2S1338, D17S1290 and D5S2500), one CODIS STR locus (D16S539) and the amelogenin locus in one reaction. Microreader™ 23sp ID system was validated according to the guidelines of "Validation Guidelines for DNA Analysis Methods (2012)" described by the Scientific Working Group on DNA Analysis Methods (SWGDAM), including PCR-based studies, sensitivity study, precision and accuracy evaluation, stutter percentage and peak height ratio, inhibitors, species specificity and DNA mixture studies. Our results suggested that Microreader™ 23sp ID system is a useful tool for identification and parentage testing.

Development of a rapid 21-plex autosomal STR typing system for forensic applications.

DNA-STR genotyping technology has been widely used in forensic investigations. Even with such success, there is a great need to reduce the analysis time. In this study, we established a new rapid 21-plex STR typing system, including 13 CODIS loci, Penta D, Penta E, D12S391, D2S1338, D6S1043, D19S433, D2S441 and Amelogenin loci.