Download Microbial DNA qPCR Assays

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Garantirana detekcija in kvantifikacija z uporabo laboratorijsko preverjenih testov
Zanesljivi rezultati z integriranimi kontrolami
Pripravljena programska orodja za preprosto interpretacijo rezultatov
Microbial DNA qPCR Assays (330025)
1X 100 µl tube Microbial DNA qPCR Assay, 1X 1.35 ml tube Microbial qPCR Mastermix
Izbor testov: Microbial DNA qPCR Assay/Assay
Izbor testovza odpornost na antibiotike: Antibiotic Resistance Gene Resource and Assay List
Microbial DNA qPCR Assay Kit (330033)
1X 20 µl tube of Microbial DNA qPCR Assay, 1X tube Positive PCR Control (20 µl), 1X tube Microbial DNA Positive
Control (50 µl), 1X tube Microbial DNA-Free Water (1.35 ml), 1X tube Microbial qPCR Mastermix (1.35 ml)
Microbial DNA qPCR Arrays (330261)
Pred pripravljeni hidrolizirani testi so naneseni na različnih PCR ploščah 96 ( pakiranje 2/12/24), 384 (pakiranje
1/4/16), 100 mestni okrogli nosilec, primernih za različne qPCR naprave. PCR miksi lahko vsebujejo ROX, Fluorescin ali
so brez dodatka. Arreji vsebujejo vse reagente za pripravo, tudi vodo brez vsebnosti mikrobne DNA.
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Testiranje Hrane: Perutnina
Testiranje Hrane: Morska hrana
Testiranje Hrane: Zelenjava
Testiranje Hrane: Mlečni izdelki
Testiranje Hrane: Meso
Vaginalna flora
Bakterijska Vaginoza
Okužbe sečil
Črevesne okužbe
Presnovne motnje
Okužbe dihal
Ustne bolezni
Sepsa
Odpornost na antibiotike
Analiza vode
Biološka obramba
Slika: Primer plošče Microbial DNA qPCR Arrays. Na plošči testiramo en vzorec z 90 Microbial testi.
Vsaka jamica vsebuje posamezen test. Vključene so tudi kontrole vzorca, pan-bakterijska, pan-glive in
pozitivna PCR kontrola
Custom Microbial DNA qPCR Arrays (330161)
Po želji izbranih 90 testov v želenem vrstnem redu pripravijo na plošče. Hidrolizirani testi so naneseni na različnih
ploščah (96(pakiranje 12/24/+12...), 384(pakiranje 6/24/+6...),100 mestni okrogli nosilec ) primernih za različne qPCR
naprave. PCR miksi lahko vsebujejo ROX, Fluorescin ali so brez dodatka. Arreji vsebujejo vse reagente za pripravo, tudi
vodo brez vsebnosti mikrobne DNA.
Minimalno naročilo je 12 (96 mestnih plošč) ali 6 (384 mestnih plošč).
Microbial DNA qPCR Multi-Assay Kits (330043)
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Kompletni kit za mikrobno profiliranje s kontrolami in reagenti
Vsebuje ploščo z experimentalno preverjenimi Microbial DNA qPCR aseji.
Uporablja se 5' hidrolizirano sondo za občutljivo in specifično detekcijo
Microbial DNA qPCR aseji, Microbial DNA Positive Control, Positive PCR Control, Microbial DNA-Free Water, and
Microbial qPCR Mastermix.
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Bartonella henselae Pathogenicity
Bordetella pertussis Pathogenicity
Brucella spp Pathogenicity
Campylobacter jejuni Pathogenicity
Chlamydia trachomatis Pathogenicity
Clostridium difficile Pathogenicity
Clostridium perfringens Pathogenicity
C. diphtheriae Pathogenicity
Enterococcus faecalis Pathogenicity
Enterohaemorrhagic E. coli Pathogenicity
Haemophilus influenzae Pathogenicity
Helicobacter pylori Pathogenicity
Legionella pneumophila Pathogenicity
Listeria monocytogenes Pathogenicity
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Mycobacterium spp Pathogenicity
Neisseria meningitidis Pathogenicity
Pseudomonas aeruginosa Pathogenicity
Salmonella enterica Pathogenicity
Shigella dysenteriae Pathogenicity
Streptococcus agalactiae Pathogenicity
Streptococcus pneumoniae Pathogenicity
Streptococcus pyogenes Pathogenicity
Vibrio cholerae Pathogenicity
Yersinia enterocolitica Pathogenicity
Yersinia spp Pathogenicity
MRSA: Methicillin-resistant S. aureus
Laboratory Rodent Testing
SHV Antibiotic Resistance Genes
Sestava Microbial DNA qPCR Multi-Assay Kita, MRSA: Methicillin-resistant S. aureus
Microbial DNA-Free Water: 338132; Microbial DNA Positive Control: 338134; Supplemental Microbial qPCR
Mastermix (Fluorescein): 330540; Supplemental Microbial qPCR Mastermix (ROX): 330530;
Tehnične informacije
Limit of detection versus lower limit of quantification.
This chart demonstrates the difference between the limit of detection
(LOD) and the lower limit of quantification (LLOQ). The LOD is
defined as the lowest concentration at which 95% of the positive
samples are detected, whereas the LLOQ is the lowest concentration
that falls within the linear range of a standard curve. LOD depends
upon the precision of the assay, and requires at least 40 replicates for
determination of a positive sample. For the Microbial DNA qPCR
Assays, LLOQ is sufficient to determine assay sensitivity.
Linearity and sensitivity of Microbial DNA qPCR Arrays.
Linearity and sensitivity for each Microbial DNA qPCR Array was determined using
synthetic templates over a 6 log serial dilution ranging from 1 copy to 1 million copies.
The following are representative results for all the qPCR assays. [A] shows the realtime amplification curves of the KPC antibiotic resistance gene qPCR assay. In [B], a
standard curve was prepared that shows that the primer efficiency equals 103%
(calculated from slope = –3.3236) and the correlation coefficient is 0.9983, indicating
optimum performance for the KPC qPCR assay. All Microbial DNA qPCR Assays
have primer efficiencies between 80–120% and correlation coefficients (R)>0.995.
The lower limit of quantification
The lower limit of quantification
(LLOQ)
(LLOQ) for antibiotic resistance gene
for all Microbial DNA qPCR
detection Microbial DNA qPCR
Assays reveals high sensitivity.
Assays reveals high sensitivity.
This chart shows the distribution of LLOQThis
for chart shows the distribution of
all Microbial DNA qPCR Assays. 93% of LLOQ
all
for Microbial DNA qPCR
Microbial DNA qPCR Assays have a LLOQ
Assays
of for antibiotic resistance gene
<100 gene copies.
detection. 95% of all antibiotic
resistance gene assays have a LLOQ
of <100 gene copies.
The lower limit of quantification
(LLOQ) for microbial identification
Microbial DNA qPCR Assays reveals
high sensitivity.
This chart shows the distribution of
LLOQ for microbial identification
Microbial DNA qPCR Assays. 92%
of all microbial identification assays
have a LLOQ of <100 gene copies.
The lower limit of quantification
(LLOQ) for virulence factor gene
detection Microbial DNA qPCR
Assays reveals high sensitivity.
This chart shows the distribution of
LLOQ for Microbial DNA qPCR
Assays for virulence factor gene
detection. 97% of all virulence factor
gene assays have a LLOQ of <100
gene copies
Microbial DNA qPCR Assays display high sensitivity even in complex metagenomic samples.
To ensure that Microbial DNA qPCR Assays performed comparably in a complex sample, where there
may be up to a thousand different microbial species, each assay was tested using stool, tooth plaque, and
sputum samples. For each sample, synthetic template targets were spiked in and the CT was compared to
synthetic template alone. PCR was performed using several sample types, which included pooled synthetic
template targets alone, stool, stool plus pooled synthetic template targets, plaque, plaque plus pooled
synthetic template targets, sputum, and sputum plus pooled synthetic template targets. If the CT<35 in
stool, plaque, or sputum samples alone, then ΔCT was calculated (i.e., CTstool + pooled synthetic template
targets – CTpooled synthetic template targets). This calculation was performed for all the assays. For each
assay, the ΔCT<3, indicating that a complex metagenomic background does not affect the performance of
each Microbial DNA qPCR Assay.
Vaginal samples positive for Gardnerella vaginalis also
show changes in commensal and bacterial vaginosis-related
microbes compared to healthy samples.
To compare any differences in the vaginal microbiome
between healthy women and women with bacterial
vaginosis, each sample that tested positive for Gardnerella
vaginalis using the Vaginal Flora Microbial DNA qPCR
Array was compared to samples from healthy women
(n=3). Fold-change in microbial species abundance was
calculated by the ΔΔCT method using human genomic
DNA to normalize. The results show that as the relative
abundance of Gardnerella vaginalis increases, the
abundance of the commensal species Lactobacillus
crispatus decreases. Also, an increase in Gardnerella
vaginalis was associated with an increase in other bacterial
vaginosis-associated microbial species. This suggests that
Lactobacillus crispatus protects the vagina from bacterial
vaginosis-associated microbial species.
Specificity of the Antibiotic Resistance Genes Microbial DNA qPCR Array is
verified by pyrosequencing.
To verify the specificity of the Antibiotic Resistance Genes Microbial DNA
qPCR Array (cat no. BAID-1901Z) results from Klebsiella pneumoniae
isolates, pyrosequencing assays were designed to detect for the presence and
sequences of SHV-156G, SHV-156D, SHV-238G240E, SHV-238S240K,
SHV-238S240E, SHV-238G240K, ermB, mefA, tetA, tetB,CTX-M-1 Group,
CTX-M-2 Group, AAC(6′)-lb-cr and aadA1. For each Klebsiella pneumoniae
isolate, results from the Antibiotic Resistance Genes Microbial DNA qPCR
Array were confirmed by pyrosequencing. Representative pyrograms for [A]
SHV-156G, [B] SHV-238/240, [C] KPC and [D] CTX-M-1 group are shown.
For SHV variants, the Antibiotic Resistance Gene Microbial DNA qPCR
Array was able to reliably distinguish single nucleotide polymorphisms
occurring at different sites.
The Vaginal Flora Microbial DNA qPCR Array provides accurate profiling
for cervical swab samples.
The vaginal microbiota is a key component influencing women’s urogenital
health. To determine what changes in the vaginal microbiota occurs during
bacterial vaginosis, the Vaginal Flora Microbial DNA qPCR Array (cat. no.
BAID-1902Z), which detects up to 90 different microbial species, was used to
test cervical swabs from healthy individuals and from patients with bacterial
vaginosis. Genomic DNA from vaginal samples originating from three
patients that tested negative for bacterial vaginosis, three patients that tested
positive for Candida, three patients that tested positive for Garderella
vaginalis, and one patient that tested positive for Trichomonas vaginalis by
BD Affirm™ VPIII Microbial Identification Test were run on the Vaginal
Flora Microbial DNA qPCR Array. Genomic DNA from ThinPrep samples
were isolated using QIAGEN’s QIAamp MinElute Media Kit and 500 ng
genomic DNA from each sample was analyzed. After the PCR run on a Roche
LightCycler 480, raw CT values were exported to the Microbial DNA qPCR
data analysis software. Positive (+) / negative (blank) / inconclusive (+/-)
results for each microbial species were determined using the identification
criteria. The results from the Vaginal Flora Microbial DNA qPCR Array were
in concordance with the BD Affirm VPIII Microbial Identification Test.
The Microbial DNA qPCR Array screens gut microbiota for the presence of antibiotic resistance genes.
The human gut microbiota is known to act as a reservoir for antibiotic resistance genes, where they can be transferred horizontally to potential pathogenic
bacteria. To detect the presence of antibiotic resistance genes from gut microbiota, stool samples from five healthy adults were collected and genomic DNA was
isolated using QIAGEN’s QIAamp DNA Stool Mini Kit. 500 ng genomic DNA from each stool sample was analyzed for presence of antibiotic resistance genes
using the Antibiotic Resistance Genes Microbial DNA qPCR Array (cat. no. BAID-1901Z). The Antibiotic Resistance Genes Microbial DNA qPCR Array
contains assays for 83 antibiotic resistance genes, assays to identify methicillin-resistant Staphylococcus aureus, and control assays. ErmB, mefA, and tetA
were found in all or most of the stool samples tested, showing that they may be highly prevalent in the gut. These antibiotic resistance genes have been reported
to be isolated from bacterial strains originating from food, suggesting a possible source of origin. This highlights the importance of increased monitoring of
antibiotic resistance reservoirs to identify potential sources of antibiotic-resistant bacteria.