• juni 18, 2021

Vergelijking van PCR en fenotypische methoden voor de detectie

Vergelijking van PCR en fenotypische methoden voor de detectie van methicillineresistente Staphylococcus aureus 

Background and goals: Resistance to methicillin in methicillin resistant strains of Staphylococcus aureus (MRSA) is because of the presence of mec-A gene, which encodes a low affinity penicillin binding protein (PBP)-2a or PBP2. Correct and fast identification of MRSA in scientific specimens is crucial for well timed determination on efficient remedy. The purpose of the research was to match three completely different strategies for detection of MRSA specifically cefoxitin disc diffusion, CHROM agar MRSA and VITEK-2 susceptibility with PCR which is the gold customary reference methodology and to seek out the antibiotic susceptibility sample of those isolates by VITEK-2.

Supplies and strategies: A Complete of 100 non-duplicate S. aureus isolates had been collected from completely different scientific samples amongst each outpatient and inpatients. Detection of MRSA amongst these isolates was accomplished by cefoxitin disc diffusion, VITEK-2, CHROM agar MRSA and PCR.

Outcomes: The sensitivity and specificity of cefoxitin disc diffusion and Vitek was discovered to be 97.2% and 100%, whereas that of CHROM agar was discovered to be 100% and 78.6%. The total prevalence of MRSA in our research by PCR was 72%.

Conclusion: Primarily based on the findings in our research, isolates which present cefoxitin zone diameter < 22 mm could be reported as MRSA. Nevertheless, these isolates which have a zone diameter between 22-24 mm, ought to ideally be confirmed by PCR.


Een snelle multiplex PCR-test voor soortidentificatie van Aziatische rijstplanthoppers (Hemiptera: Delphacidae) en de toepassing ervan op nimfen in het vroege stadium in rijstvelden

Rice (Oryza sativa L.) is the principle cereal crop in lots of Asian nations. The Asian rice planthoppers, Nilaparvata lugens (Stål) (brown planthopper), Sogatella furcifera (Horváth) (white-backed planthopper), and Laodelphax striatellus (Fallén) (small brown planthopper) (Hemiptera: Delphacidae), are essentially the most economically vital pests of rice. These three rice planthopper species usually co-occur in the identical paddy area. Historically, species identification of people of the three rice planthopper species has relied on morphological characters, however correct discrimination of early-instar nymphs may be very tough, even for professional researchers. On this research, we developed a fast one-step multiplex PCR assay utilizing conserved and species-specific 5.8S-ITS2 rDNA gene primers for simultaneous identification of people of the three rice planthopper species.

The multiplex PCR outcomes confirmed that the three rice planthopper species may very well be recognized precisely based mostly on the size of the resultant amplicon, whatever the particular person developmental stage. Moreover, we utilized this assay for the primary correct quantification of early-instar nymphs of every rice planthopper species in paddy fields. Notably, we discovered that the species composition of early-instar nymphs can’t be extrapolated from that of adults. Thus, the multiplex PCR assay developed right here facilitates detection of every rice planthopper species firstly of outbreaks in paddy fields.

Retrospectieve observationele RT-PCR-analyses van 688 child’s van 843 SARS-CoV-2-positieve moeders, placenta-analyses en diagnostische analyses, beperkingen suggereren dat verticale transmissie mogelijk is

Analysis query: Is there vertical transmission (from mom to child antenatally or intrapartum) after SARS-CoV-2 (COVID-19) contaminated being pregnant?

Research design: A scientific search associated to SARS-CoV-2 (COVID-19), being pregnant, neonatal issues, viral and vertical transmission. The length was from December 2019 to Might 2020.

Outcomes: A complete of 84 research with 862 COVID optimistic girls had been included. Two research had ongoing pregnancies whereas 82 research included 705 infants, 1 miscarriage and 1 medical termination of being pregnant (MTOP). Most publications (50/84, 59.5%), reported small numbers (<5) of optimistic infants. From 75 research, 18 infants had been COVID-19 optimistic. The primary reverse transcription polymerase chain response (RT-PCR) diagnostic take a look at was accomplished in 449 infants and a couple of losses, 2nd RT-PCR was accomplished in 82 infants, IgM exams had been accomplished in 28 infants, and IgG exams had been accomplished in 28 infants. On the primary RT-PCR, 47 research reported time of testing whereas 28 research didn’t. Optimistic leads to the primary RT-PCR had been seen in 14 infants. Earliest examined at beginning and the typical time of the consequence was 22 hours. Three infants with unfavorable first RT-PCR grew to become optimistic on the second RT-PCR at day 6, day 7 and at 24 hours which continued to be optimistic at 1 week.4 research with a complete of Four placental swabs had been optimistic demonstrating SARS-CoV-2 localised within the placenta. In 2 research, 10 exams for amniotic fluid had been optimistic for SARS-CoV-2. These 2 infants had been discovered to be optimistic on RT-PCR on serial testing.

Conclusion: Diagnostic testing mixed with incubation interval and placental pathology point out a robust chance that intrapartum vertical transmission of SARS-CoV-2 (COVID-19) from mom to child is feasible.

Gevoeligheid van SARS-CoV-2 RNA-polymerasekettingreactie met behulp van een klinische en radiologische referentiestandaard: Klinische gevoeligheid van SARS-CoV-2 PCR

Goals: Diagnostic exams for SARS-CoV-2 are vital for epidemiology, scientific administration, and an infection management. Limitations of oro-nasopharyngeal real-time PCR sensitivity have been described based mostly on comparisons of single exams with repeated sampling. We assessed SARS-CoV-2 PCR scientific sensitivity utilizing a scientific and radiological reference customary.

Strategies: Between March-Might 2020, 2060 sufferers underwent thoracic imaging and SARS-CoV-2 PCR testing. Imaging was independently double- or triple-reported (if discordance) by blinded radiologists in line with radiological standards for COVID-19. We excluded asymptomatic sufferers and people with various diagnoses that would clarify imaging findings. Associations with PCR-positivity had been assessed with binomial logistic regression.

Outcomes: 901 sufferers had attainable/possible imaging options and scientific signs of COVID-19 and 429 sufferers met the scientific and radiological reference case definition. SARS-CoV-2 PCR sensitivity was 68% (95% confidence interval 64-73), was highest 7-Eight days after symptom onset (78% (68-88)) and was decrease amongst present people who smoke (adjusted odds ratio 0.23 (0.12-0.42) p<0.001).

Conclusions: In sufferers with scientific and imaging options of COVID-19, PCR take a look at sensitivity was 68%, and was decrease amongst people who smoke; a discovering that would clarify observations of decrease illness incidence and that warrants additional validation. PCR exams ought to be interpreted contemplating imaging, symptom length and smoking standing.

Vergelijking van microscopie en PCR voor detectie van Giardia Lamblia en Entamoeba Histolytica in monsters van menselijke ontlasting in een omgeving met beperkte middelen in West-Kenia

Background: Correct prognosis of Giardia lamblia and Entamoeba histolytica is vital since these intestinal parasites account for a big proportion of morbidity and mortality globally. Microscopy is the important thing diagnostic take a look at used for prognosis of the 2 parasites. Different exams together with fast diagnostic exams and polymerase chain response have been developed to enhance the detection of those parasites. Most of those newer exams are usually not inexpensive in useful resource restricted settings, therefore the over reliance on microscopy. The target of this research was to find out the reliability of microscopy in a useful resource restricted setting in Western Kenya, a area endemic for the 2 intestinal parasites.

Strategies: Polymerase chain response, the gold customary take a look at, was carried out on stool samples suspected for G. lamblia and E. histolytica. Microscopy was then carried out on the identical samples and the 2 exams in contrast.

Outcomes: Microscopy was discovered to be 64.4% delicate, 86.6% particular for the detection of G. lamblia. Moreover, this take a look at was 64.2% delicate and 83.6% particular for the prognosis of E. histolytica. Cohen’s kappa values of 0.51 and 0.47 had been decided for microscopy for G. lamblia and E. histolytica respectively. McNemar’s take a look at revealed a vital distinction between the 2 exams, P<0.001.

Conclusion: This research discovered microscopy to be a dependable diagnostic take a look at on this useful resource restricted setting.

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