Control Primer Research Articles

Pair-wise comparison analysis of differential expression of mRNAs in early and advanced stage primary colorectal adenocarcinomas

ObjectivesTo characterise the mRNA expression patterns of early and advanced stage colorectal adenocarcinomas of Malaysian patients.DesignComparative expression analysis.Setting and participantsWe performed a combination of annealing control primer (ACP)-based PCR and...

Efficacy of a fluoride-releasing orthodontic primer in reducing demineralization around brackets: An in-vivo study

A new, highly filled primer is currently marketed as a fluoride delivery system effective in reducing white spot lesions in orthodontic patients. However, no studies in the literature support this claim. The purpose of this in-vivo study was to investigate the retention and the efficacy of this primer in reducing the formation of white spot lesions. In each patient for whom premolar extractions were planned (n = 22), 1 premolar was randomly chosen as the experimental tooth for the application of the fluoride delivery system (Opal Seal; Ultradent Products, South Jordan, Utah), and the contralateral tooth was assigned as the control to receive the standard treatment (Transbond XT; 3M Unitek, Monrovia, Calif). After the bonding procedures, separators were placed around the premolar brackets to encourage plaque retention over 8 weeks. After the extractions, the tooth surfaces were evaluated visually and with microhardness techniques for demineralization. Primer retention was also investigated. There were no statistically significant differences in the numbers of white spot lesions between the 2 groups. The primer retention was calculated as 50%. The results indicated no significant difference between the efficacies of the fluoride-releasing primer and the control primer in reducing demineralization over the duration of the study.

염과 건조 스트레스 조건에서 톨 페스큐의 종자 발아율과 유전자 발현 변화분석

ABSTRACT Salinity and drought stresses are probably the most significant abiotic factor limiting plant’s growth, also negatively affect seed germination and early seedling development. To study on effect of NaCl and PEG stress on seed germination and gene expression pattern of tall fescue, the levels of NaCl and PEG-induced water stresses were determined in first experiment. Different concentration of NaCl (0 to 350 mM) and PEG (0 to 30%) were used for seed treatment. Seed Germination percentage reduced with increasing osmotic potential of growth medium either due to NaCl or PEG. Seeds were not germinate at 350 mM NaCl or 30% PEG treatment. On the basis of the results, Kentucky31(E-) had more resistant than Fawn in both stress conditions. Furthermore, we have used an annealing control primer-based differential display reverse transcription-polymerase chain reaction method to identify salt- and drought stress-induced differentially expressed genes (DEGs) in tall fescue leaves. Using 120 annealing control primers, a total of 4 genes were identified and sequenced. The possible roles of the identified DEGs are discussed in the context of their putative role during salinity and drought stresses.(

P38δ Regulates p53 to Control p21Cip1 Expression in Human Epidermal Keratinocytes

PKCδ suppresses keratinocyte proliferation via a mechanism that involves increased expression of p21(Cip1). However, the signaling mechanism that mediates this regulation is not well understood. Our present studies suggest that PKCδ activates p38δ leading to increased p21(Cip1) promoter activity and p21(Cip1) mRNA/protein expression. We further show that exogenously expressed p38δ increases p21(Cip1) mRNA and protein and that p38δ knockdown or expression of dominant-negative p38 attenuates this increase. Moreover, p53 is an intermediary in this regulation, as p38δ expression increases p53 mRNA, protein, and promoter activity, and p53 knockdown attenuates the activation. We demonstrate a direct interaction of p38δ with PKCδ and MEK3 and show that exogenous agents that suppress keratinocyte proliferation activate this pathway. We confirm the importance of this regulation using a stratified epidermal equivalent model, which mimics in vivo-like keratinocyte differentiation. In this model, PKCδ or p38δ knockdown results in reduced p53 and p21(Cip1) levels and enhanced cell proliferation. We propose that PKCδ activates a MEKK1/MEK3/p38δ MAPK cascade to increase p53 levels and p53 drives p21(Cip1) gene expression.

Establishment of stable SGC-7901 gastric cancer cell line with cortactin down-regulation

Objective To establish a stable SGC-7901 gastric cancer cell line with cortactin downregulation.Methods Three pairs of primers specific to knockdown cortactin and a pair of control primers were designed and synthesized.The primers were annealed and inserted into PLKO.1 plasmids.The plasmids,psPAX2,and PCMV-VSVG were transfected into HEK293T cells simultaneously,and then lentiviral particles were produced and collected.The lentiviral particles were used to infect SGC-7901 cells and puromycin was used to select stable cell lines.Three stable cell lines were selected at last.Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were conducted to detect the expression level of cortactin mRNA and protein respectively.Results The expression levels of cortactin mRNA and protein were significantly down-regulated.The mRNA and protein levels of cortactin in PLKO.1 + short hairpin RNA (shRNA)3 cell line were significantly decreased,about 10％ of the control stable cell line.Conclusion The stable SGC-7901 gastric cell lines with cortactin down-regulation were established successfully. Key words: Cortactin; Short hairpin RNA ; Gastric

The identification and detection of a unique gene in Salmonella enterica subsp. enterica Weltevreden using PCR-based approach

IntroductionSalmonella enterica are important food-borne pathogens that cause human gastroenteritis, bacteremia and subsequent focal infections. The incidences of food-borne salmonellosis due to non-typhoidal Salmonella serotypes are increasing periodically in Malaysia. Other than the two predominant non-typhoidal Salmonella serotypes, S. Typhimurium and S. Enteritidis, an emerging serotype, S. Weltevreden, is increasingly being isolated from hospitalized-gastroenteritis patients in Malaysia. Serotyping is the conventional method for differentiation of this pathogen from the 2500 other serotypes known to exist. However, this method has a number of disadvantages including low specificity and high cost. Therefore, a molecular method for identification of S. Weltevreden in clinical isolates based on amplification of a specific DNA sequence present in its genome is proposed. MethodsA total of 234 clinical isolates were collected from the Department of Clinical Microbiology and Parasitology, HUSM from year 2010-2012. There were 49 S. Weltevreden isolates which have been confirmed by the Institute of Medical Research (IMR) using conventional serotyping method. Two sets of primers targeting a unique gene in S. Weltevreden (SWE349) and a pan-Salmonella gene (invA284) were designed using Primer3 online software, and a multiplex-PCR (mPCR) assay was developed to detect S. Weltevreden in clinical isolates. The optimized S. Weltevreden mPCR assay was evaluated on a panel of known Salmonella and non-Salmonella spp. to determine the specificity and sensitivity of the mPCR assay. Results & DiscussionThe S. Weltevreden serovar specific primer, SWE349, and the internal control primer, invA284, resulted in 2 PCR products of 349bp and 284bp, respectively. The mPCR assay for S. Weltevreden targeting gene SWE349 showed 100% sensitivity (49/49) and specificity (36/36) in concordance with the conventional serotyping method in detecting only S. Weltevreden clinical strains, but not from the panel of 26 non-typhoidal Salmonella and 10 non- Salmonella isolates. ConclusionA cost effective, sensitive and specific mPCR assay has been developed for the detection and confirmation of Salmonella Weltevreden from clinical isolates of gastroenteritis patients, using genes SWE349 and invA284.

Candida albicans aspects of novel silane system–coated titanium and zirconia implant surfaces

The aim of the present study was to evaluate the effect of novel silane system coatings on zirconia and titanium implant surfaces and the attachment of the fungal pathogen Candida albicans. Titanium and zirconia specimens were silica-coated and silanized either with a commercial silane primer (RelyX Ceramic Primer™, 3M ESPE) or a novel silane system primer. The novel silane system primer was a blend of 1.0 vol% 3-acryloxypropyltrimethoxysilane and 0.3 vol% bis-1,2-(triethoxysilyl)ethane diluted in acidified ethanol-water solvent. The surface roughness (Ra ), the surface free energy and the chemical composition of substrate surfaces after treatments were evaluated. C. albcans biofilms were developed on silica-coated + silanized surfaces during 48 h of incubation time. Colony forming units (CFU) and real-time PCR (RT-PCR) quantified the cells on the material surfaces. Statistical analyses were carried out by 1-way ANOVA, Tukey post hoc and Games Howell post hoc test at 5% significance level (p). On zirconia and titanium surfaces, the Ra and the chemical composition of the specimens were equal (P < 0.05). The surface free energy was decreased on titanium specimens and increased on zirconia specimen after silanization. CFU of C. albicans was significantly lower on zirconia coated with RelyX Ceramic Primer™, (P < 0.001) and on titanium coated with both silanes (P = 0.002). RT-PCR revealed no differences between the mean quantities of C. albicans (P ≥ 0.067). Silica-coating and silanization had modified the titanium and zirconia surfaces significantly. Both the control and experimental silane primers might inhibit the biofilm formation of C. albicans.

Isolation of differentially expressed genes related to salt tolerance in peanut

Salinity is one of the most serious environmental factors influencing crop growth and development, especially during germination period. In peanut, screening for salinity tolerance has been reported by several investigators, but there are few reports regarding isolation and identification of salt responsive genes from salt tolerant genotypes. In this study, we compared gene expression profile of a large-seeded saline tolerant peanut line 10L19 imbibed in 1.0% sodium chloride solution and seeds of the same genotype treated with distilled water, using the annealing control primer (ACP)-based GeneFishing technology. Total RNAs were extracted from seeds sampled at 1, 6 and 24 h after imbibition initiation. With 40 ACPs, a total of 20 differentially expressed transcripts were isolated. Thirteen of them were found to have high homology to genes with known functions in GenBank nr database. BLAST search revealed that those genes were related to gene expression regulation, signalling transduction, development and stress tolerance, among which S-38 was of direct relevance to hyperosmotic salinity response. S-14, S-15, S-17, S-28, S-31, S-32 and S-38, encoding ethylene-responsive element binding factor 4, DNA-directed RNA polymerase subunit, oleosin 5, polygalacturonase-inhibiting protein, vacuolarprocessing enzyme precursor, ribosomal protein s7 and nac domain protein, respectively, were selected and their relative expression levels determined by qRT-PCR using β-actin as internal control. The results confirmed that all the seven genes were expressed differentially in stressed and control peanut seeds. The highest relative expression for S-17 and S-38 was both at 6 h, while that for the rest genes was at 24 h.

A simple and efficient method to isolate LTR sequences of plant retrotransposon.

Retrotransposons (RTNs) have important roles in the formation of plant genome size, structure, and evolution. Ubiquitous distributions, abundant copy numbers, high heterogeneities, and insertional polymorphisms of RTNs have made them as excellent sources for molecular markers development. However, the wide application of RTNs-based molecular markers is restricted by the scarcity of the LTR (long terminal repeat) sequences information. A new, simple, and efficient method to isolate LTR sequences of RTNs was presented based on the degenerate RNase H nested primers and PPT (polypurine tract) primer of RTNs in tree peony. This method combined the characteristics and advantages of high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR), annealing control primer (ACP) system, and suppression PCR method. Nineteen LTR sequences were isolated using this new method in tree peony and the applicability of the LTR sequences based markers was validated by further SSAP analysis. The results showed that the new method is simple, of low-cost, and highly efficient, which is just conducted by three rounds of PCR and does not need any restriction enzymes and adapters, much less the hybridizations. This new method is rapid, economical, and cost- and time-saving, which could be easily used to isolate LTR sequences of RTNs.

Innovative self-etching dental adhesives formulated using reactive nanogel additives

Purpose: Enzymatic degradation of poorly resin-infiltrated collagen and hydrolysis of the resin-based polymeric matrix are interfacial degradation mechanisms. However, since the depth of demineralization and resin infiltration in self-etching adhesives (SEAs) occurs simultaneously, the hydrolysis of the hydrophilicmonomers requires further investigation. The aim of this study was to evaluate the microtensile bond strength (mTBS) and the confocal interfacial ultra-morphology of innovative nanogel-modified two-step SEAs bonded to dentin. Methods and materials: Nanogel copolymers were synthesized using isobornyl-methacrylate, urethane-dimethacrylate (UDMA, Esstech, USA)in presence of 2-Mercaptoethanol as a chain-transfer agent and photo-polymerized (2h) in toluene solution with bis(2,4,6-trimethylbenzoyl)phenylphosphine oxide (320-500nm, 2500 mW/cm(2)). A two-step self-etching adhesive was prepared using UDMA, 2-hydroxyethyl methacrylate and 10-methacryloxydecyldihydrogen-phosphate (40/25/15wt%). Camphorquinone and N,N-ethyl-dimethylaminobenzoate were added (0.3 and 0.7 wt%, respectively). Ethanol (14wt%) and water (5wt%) were also included to solvate the resin blend to create a control primer, while non-solvated resin was used as the adhesive. The nanogels were added to create an experimental primer (15wt%) and adhesive (25wt%). Clearfil SE-Bond (CSE; Kuraray, Japan) was also used as control SEA. Several bonding primer/adhesive combinations were applied on middle dentin (5 specimens each group), light cured (20s) and resin composite (N’Durance, Septodont, USA) build-ups (5mm) were created and light-cured (40s each 1-mm layer). The resin-bonded specimens beams (0.9mm(2)) were submitted to mTBS after 24h or 6 months of water aging. The results were statistically analysed by two-way ANOVA and Student–Newman–Keuls test used (p 0.05) after 24h. However, only the control nanogel-free primer/adhesive and the primer/adhesive both containing nanogels showed significant mTBS reduction (p<0.05) after 6 months. The greatest resistance to water aging was attained when the nanogels where present only in the adhesive. The CLSM analysis showed nanogels infiltration within the hybrid layer and low micropermeability both after 24h and 3 months of water aging. Conclusion: The use of nanogels within the composition of two-step SEAs improved their long-termdurability due to infiltration within the hybrid layer, so making the resin-dentine interface hydrophobic andmore resistant to hydrolytic degradation.

Genus‐specific PCR assay for screening Arcobacter spp. in chicken meat

The number of emerging pathogenic species described within the genus Arcobacter has increased rapidly during the last few years. In this work a genus-specific polymerase chain reaction (PCR) assay was developed for detection of the species of Arcobacter most commonly associated with foods. The assay uses primers designed to amplify an 85 bp DNA fragment on the 16S rRNA gene and was applied to the detection of Arcobacter spp. in retail chicken meat. Primer specificity was tested against a panel of Arcobacter spp., related Campylobacter and Helicobacter spp. and other food bacteria. Arcobacter primers consistently and selectively amplified the expected DNA fragment in all tested Arcobacter spp. Bacterial control primers confirmed the presence of amplifiable DNA in the samples. The applicability of the PCR assay to food was validated through screening of fresh retail chicken samples for the presence of Arcobacter spp., with a result of 45% (23 out of 51) positive samples. The genus-specific PCR assay developed has the potential to be used as a quick and sensitive alternative method for the survey of Arcobacter contamination in meats.

Red Cell Distribution Width Identifies Cerebral Venous Sinus Thrombosis in Patients With Headache

Diagnosing cerebral venous sinus thrombosis (CVST) in patients referring to emergency service or neurology outpatient unit with complaints of headache is a challenging task. Magnetic resonance (MR) venography is the gold standard, but there are limitations regarding its use. To evaluate the validity of red cell distribution width (RDW) in CVST diagnosis in patients presenting with headache. A total of 138 patients comprising 37 patients with CVST and 101 control primer headache cases were included in this retrospective cross-sectional study. Control group consists of hospitalized patients with primary headache. Venous blood hemoglobin (Hb), platelet, mean corpuscular volume (MCV), RDW, fibrinogen, and vitamin B12 levels of the patients were recorded at the first referral. Diagnosis of CVST was established by MR venography. The RDW ratio of patients with a diagnosis of CVST was significantly higher than that in patients with primary headache (15.3 ± 1.4 vs 13.3 ± 0.5; P <.0001). Fibrinogen and vitamin B12 levels were not significantly different between the 2 groups. In a total of 11 patients, there was more than 1 thrombosis. In 21 of the patients with CVST, gene mutation was detected. There was no significant difference between the patients with and without mutations regarding RDW values. Diagnostic validity of RDW was found to be excellent in differentiating patients with CVST and primary headache (area under the curve = 0.996; 95% CI: 0.990-1.000). Optimum RDW cutoff value was determined as 14.1% (sensitivity: 91.9%, specificity: 99%, positive predictive value: 92.8, negative predictive value: 0.082). We suppose that among patients presenting with the complaint of headache, RDW value may lead to diagnose CVST.

Microarrays for high‐throughput genotyping ofMICAalleles using allele‐specific primer extension

The role of major histocompatibility complex (MHC) class I chain-related gene A (MICA), a ligand of NKG2D, has been defined in human diseases by its allele associations with various autoimmune diseases, hematopoietic stem cell transplantation (HSCT) and cancer. This study describes a practical system to develop MICA genotyping by allele-specific primer extension (ASPE) on microarrays. From the results of 20 control primers, strict and reliable cut-off values of more than 30,000 mean fluorescence intensity (MFI) as positive and less than 3000 MFI as negative, were applied to select high-quality specific extension primers. Among 55 allele-specific primers, 44 primers could be initially selected as optimal primer. Through adjusting the length, six primers were improved. The other failed five primers were corrected by refractory modification. MICA genotypes by ASPE on microarrays showed the same results as those by nucleotide sequencing. On the basis of these results, ASPE on microarrays may provide high-throughput genotyping for MICA alleles for population studies, disease-gene associations and HSCT.

Reverse Transcription Polymerase Chain Reaction-based System for Simultaneous Detection of Multiple Lily-infecting Viruses.

A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon province of Korea and used to evaluate the detection system. Simplex and multiplex RT-PCR were performed using virus-specific primers to detect single-or mixed viral infections in lily plants. Our results demonstrate the selective detection of 3 different viruses (CMV, LMoV and LSV) by using specific primers as well as the potential of simultaneously detecting 2 or 3 different viruses in lily plants with mixed infections. Three sets of primers for each target virus, and one set of internal control primers were used to evaluate the detection system for efficiency, reliability, and reproducibility.

Application of novel loop-mediated isothermal amplification (LAMP) for rapid authentication of the herbal tea ingredient Hedyotis diffusa Willd.

Hedyotis diffusa Willd. (Baihuasheshecao) is an ingredient of herbal teas commonly consumed in the Orient and tropical Asia for cancer treatment and health maintenance. In the market, this ingredient is frequently adulterated by the related species Hedyotis corymbosa (L.) Lam. The objective of this study is to develop a novel loop-mediated isothermal amplification (LAMP) technique to differentiate H. diffusa from its adulterant H. corymbosa. A set of four internal control primers (F3, FIP, BIP and B3) were designed based on six loci in the internal transcribed spacer (ITS) for LAMP of both H. diffusa and H. corymbosa. Two specific primers (S_F3 and S_FIP) were designed for specific LAMP detection of H. diffusa only. Our data showed that LAMP was successful for both H. diffusa and H. corymbosa in internal control. In contrast, only H. diffusa was detected in specific LAMP using the specific primers S_F3 and S_FIP. This study showed that LAMP was useful to differentiate H. diffusa from its adulterant H. corymbosa. This study is significant for the verification of the authenticity for better quality control of this common herbal tea ingredient. The strategy of including an internal control assures the quality of the concerned DNA region for LAMP.

Identification and Expression of Skinning Injury-responsive Genes in Sweetpotato

Skinning injury in sweetpotatoes (Ipomoea batatas) is responsible for significant postharvest loss resulting from storage diseases and weight loss. Unfortunately, there is no report on the genes involved in wound healing of sweetpotato and a better understanding will facilitate improved breeding strategies. An annealing control primer (ACP) system was used to identify genes expressed after skinning injury of sweetpotato cultivar LA 07-146 storage roots. Using 20 ACPs, 63 differentially expressed genes (DEGs) were identified. Functional annotation of the DEGs revealed that genes previously shown to respond to dehydration, those involved in wounding response, and the lignin and suberin biosynthesis pathways were induced in response to skinning. Expression analysis of 18 DEGs through quantitative reverse transcription–polymerase chain reaction (PCR) showed that DEGs involved in lignin and suberin pathways were up-regulated after 8 and 12 hours of skinning. Other genes showed up- or down-regulation in their transcript abundance depending on the time the storage root was sampled after intentional skinning. The genes up-regulated in response to skinning may be useful to identify expression markers for screening sweetpotato lines tolerant to skinning injury in breeding programs.

Identification of Differentially Expressed Genes by Gabapentin in Cultured Dorsal Root Ganglion in a Rat Neuropathic Pain Model

Neuropathic pain is a chronic pain disorder caused by nervous system lesions as a direct consequence of a lesion or by disease of the portions of the nervous system that normally signal pain. The spinal nerve ligation (SNL) model in rats that reflect some components of clinical pain have played a crucial role in the understanding of neuropathic pain. To investigate the direct effects of gabapentin on differential gene expression in cultured dorsal root ganglion (DRG) cells of SNL model rats, we performed a differential display reverse transcription-polymerase chain reaction analysis with random priming approach using annealing control primer. Genes encoding metallothionein 1a, transforming growth factor-β1 and palmitoyl-protein thioesterase-2 were up-regulated in gabapentin-treated DRG cells of SNL model rats. The functional roles of these differentially expressed genes were previously suggested as neuroprotective genes. Further study of these genes is expected to reveal potential targets of gabapentin.

Effects of dual antibacterial agents MDPB and nano-silver in primer on microcosm biofilm, cytotoxicity and dentine bond properties

ObjectivesThe objective of this study was to investigate the effects of dentine primer containing dual antibacterial agents, namely, 12-methacryloyloxydodecylpyridinium bromide (MDPB) and nanoparticles of silver (NAg), on dentine bond strength, dental plaque microcosm biofilm response, and fibroblast cytotoxicity for the first time. MethodsScotchbond Multi-Purpose (SBMP) was used as the parent bonding agent. Four primers were tested: SBMP primer control (referred to as “P”), P+5% MDPB, P+0.05% NAg, and P+5% MDPB+0.05% NAg. Dentine shear bond strengths were measured using extracted human teeth. Biofilms from the mixed saliva of 10 donors were cultured to investigate metabolic activity, colony-forming units (CFU), and lactic acid production. Human fibroblast cytotoxicity of the four primers was tested in vitro. ResultsIncorporating MDPB and NAg into primer did not reduce dentine bond strength compared to control (p>0.1). SEM revealed well-bonded adhesive–dentine interfaces with numerous resin tags. MDPB or NAg each greatly reduced biofilm viability and acid production, compared to control. Dual agents MDPB+NAg had a much stronger effect than either agent alone (p<0.05), increasing inhibition zone size and reducing metabolic activity, CFU and lactic acid by an order of magnitude, compared to control. There was no difference in cytotoxicity between commercial control and antibacterial primers (p>0.1). ConclusionsThe method of using dual agents MDPB+NAg in the primer yielded potent antibacterial properties. Hence, this method may be promising to combat residual bacteria in tooth cavity and invading bacteria at the margins. The dual agents MDPB+NAg may have wide applicability to other adhesives, composites, sealants and cements to inhibit biofilms and caries.

Identification of two novel female-specific DNA sequences in half-smooth tongue sole, Cynoglossus semilaevis

Half-smooth tongue sole (Cynoglossus semilaevis) is a kind of cultured teleost fish with larger size of females than males. In order to produce all-female progeny, it is necessary to distinguish the genetic females and males precisely. In this study, a W chromosome library, which had been constructed by laser microdissection of W chromosomes, was screened. Among four hundreds and twelve pairs of designed PCR primers, two pairs (W045 and W131) could produce female-specific fragments in fifty individuals. To verify the primers and optimize the method, another one hundred individuals were tested by multiplex PCR with both W045 and a pair of positive control primers. Furthermore, two sex-reversed individuals were also identified correctly. Fluorescent in situ hybridization (FISH) further confirmed that fragments form W045 and W131 primers only localized to the W chromosome in female individuals, and the FISH technique with the two female-specific fragments could help to isolate super-female individuals. The identification of two novel female-specific DNA sequences and the techniques used here may be applied to distinguish the genetic gender of half-smooth tongue sole precisely. Besides, these also help us to obtain a better understanding of fish sex determination.

Rapid and Inexpensive Screening of Genomic Copy Number Variations Using a Novel Quantitative Fluorescent PCR Method

Detection of human microdeletion and microduplication syndromes poses significant burden on public healthcare systems in developing countries. With genome-wide diagnostic assays frequently inaccessible, targeted low-cost PCR-based approaches are preferred. However, their reproducibility depends on equally efficient amplification using a number of target and control primers. To address this, the recently described technique called Microdeletion/Microduplication Quantitative Fluorescent PCR (MQF-PCR) was shown to reliably detect four human syndromes by quantifying DNA amplification in an internally controlled PCR reaction. Here, we confirm its utility in the detection of eight human microdeletion syndromes, including the more common WAGR, Smith-Magenis, and Potocki-Lupski syndromes with 100% sensitivity and 100% specificity. We present selection, design, and performance evaluation of detection primers using variety of approaches. We conclude that MQF-PCR is an easily adaptable method for detection of human pathological chromosomal aberrations.