scholarly journals Rapid detection of SARS-CoV-2 infection by multicapillary column coupled ion mobility spectrometry (MCC-IMS) of breath. A proof of concept study

2020 ◽  
Author(s):  
Claus Steppert ◽  
Isabel Steppert ◽  
William Sterlacci ◽  
Thomas Bollinger
Keyword(s):  
Ion Mobility ◽  
Influenza A ◽  
Viral Disease ◽  
Pcr Analysis ◽  
Proof Of Concept ◽  
Influenza Epidemic ◽  
Rt Pcr ◽  
Respective Group ◽  
Non Invasive ◽  
Multicapillary Column

Abstract There is an urgent need for screening of patients having a communicable viral disease to cut infection chains. We could recently demonstrate that MCC-IMS of breath is able to identify Influenza-A infected patients. With decreasing Influenza epidemic and upcoming SARS-CoV-2 infections we went on and also analysed patients with suspected SARS-CoV-2 infections.75 patients, 34m, 41f, aged 64.4 ± 15.4 years, 14 positive for Influenza-A, 16 positive for SARS-CoV-2, the remaining 44 patients were used as controls. In one patient RT-PCR was highly suspicious of SARS-CoV-2 but initially inconclusiveBesides RT-PCR analysis of nasopharyngeal swabs all patients underwent MCC-IMS analysis of breath. There was no difference in gender or age according to the groups.97.3% of the patients could be correctly classified to the respective group by discriminant analysis. Even the inconclusive patient could be mapped to the SARS-CoV-2 group applying the discrimination function.ConclusionMCC-IMS is able to detect SARS-CoV-2 infection and Influenza-A infection in breath. As this method provides exact, fast non-invasive diagnosis it should be further developed for screening of communicable viral diseases.Trial registrationClinicalTrial.gov, NCT04282135 Registered 20 February 2020 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT04282135?term=IMS&draw=2&rank=1

scholarly journals Rapid detection of SARS-CoV-2 infection by multicapillary column coupled ion mobility spectrometry (MCC-IMS) of breath. A proof of concept study

2020 ◽  
Author(s):  
Claus Steppert ◽  
Isabel Steppert ◽  
Gunther Becher ◽  
William Sterlacci ◽  
Thomas Bollinger
Keyword(s):  
Discriminant Analysis ◽  
Ion Mobility ◽  
Influenza A ◽  
Cross Validation ◽  
Viral Disease ◽  
Pcr Analysis ◽  
Proof Of Concept ◽  
Influenza Epidemic ◽  
Rt Pcr ◽  
Non Invasive

AbstractThere is an urgent need for screening patients of having a communicable viral disease to cut infection chains.We could recently demonstrate that MCC-IMS of breath is able to identify Influenza-A infected patients. With decreasing Influenza epidemic and upcoming SARS-CoV-2 infections we extended our study to the analysis of patients with suspected SARS-CoV-2 infections.51 patients, 23m, 28f, aged 64 ± 16 years, were included in this study.Besides RT-PCR analysis of nasopharyngeal swabs all patients underwent MCC-IMS analysis of breath. 16 patients, 7m, 9f, were positive for SARS-CoV-2 by RT-PCR. There was no difference in gender or age according to the groups.Stepwise canonical discriminant analysis could correctly classify the infected and non-infected subjects in 98% by cross-validation. Afterwards we combined the Influenza-A sub study and the SARS-CoV-2-sub study for a total of 75 patients, 34m, 41f, aged 64.8 ± 1.8 years, 14 positive for Influenza-A, 16 positive for SARS-CoV-2, the remaining 44 patients were used as controls. In one patient RT-PCR was highly suspicious of SARS-CoV-2 but inconclusive.There was no imbalance between the groups for age or gender.97.3% of the patients could be correctly classified to the respective group by discriminant analysis. Even the inconclusive patient could be mapped to the SARS-CoV-2 group applying the discrimination function.ConclusionMCC-IMS is able to detect SARS-CoV-2 infection and Influenza-A infection in breath. As this method provides exact, fast non-invasive diagnosis it should be further developed for screening of communicable viral diseases.Study registration: NCT04282135

scholarly journals Rapid non-invasive detection of Influenza-A-infection by multicapillary column coupled ion mobility spectrometry

2020 ◽  
Vol 15 (1) ◽  
pp. 011001
Author(s):  
Claus Steppert ◽  
Isabel Steppert ◽  
Thomas Bollinger ◽  
William Sterlacci
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Influenza A ◽  
Non Invasive ◽  
Multicapillary Column

scholarly journals Artemia sp. as a DNA vaccine vector for common carp Cyprinus carpio larvae

2014 ◽  
Vol 12 (1) ◽  
pp. 54
Author(s):  
Sri Nuryati ◽  
Sekar Sulistyaning Hadiwibowo ◽  
. Alimuddin
Keyword(s):  
Dna Vaccine ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Herpes Virus ◽  
Oral Vaccine ◽  
Viral Disease ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Glycoprotein Gene ◽  
Herpès Virus

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle" align="center"> </p><p class="NoParagraphStyle">Koi herpes virus (KHV) is one of the most common impetuses for disease on common carp <em>Cyprinus carpio</em>. Generally, viral disease is difficult to cure because virus is intra-cellular parasite, that virus survives, multiplies, and lives only if it on the host cell. Oral vaccine delivery through <em>Artemia</em> sp. is of one alternative way to overcome this problem. This experiment was carried out by analysis DNA vaccine expression encoding of glycoprotein gene (GP-11) on <em>C. carpio</em>. Bacteria containing plasmid Krt-GP-11 as vaccine is served through <em>Artemia </em>sp. as a vector. <em>Artemia</em> sp. was given for one and two times a week to three weeks old common carp. Organs of fish fed by <em>Artemia</em> sp. were analyzed every three days after vaccination. The expression of GP-11 in kidney in each treatment is also observed by the use of RT-PCR method, within ten days after vaccination. The experiment showed that dose of DNA vaccine in whole bacteria could be expressed is 10<sup>6</sup> cfu/mL in a once or twice provisions a week. DNA vaccine could be detected in three organs. RT-PCR analysis also showed that the expression of GP-11 can be detected in all tested organs. In conclusion, <em>Artemia</em> sp. can be used as a vector to carry plasmid GP-11 vaccine for common carp <em>Cyprinus carpio</em> larvae.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: DNA vaccine, KHV, <em>Artemia</em> sp., common carp</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle" align="center"><strong>ABSTRAK</strong></p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Salah satu penyakit pada ikan mas (<em>Cyprinus carpio</em>) yang disebabkan oleh virus adalah <em>koi herpes virus</em> (KHV). Penyakit yang disebabkan oleh virus umumnya sulit untuk disembuhkan karena virus merupakan parasit intraseluler, yaitu virus hanya dapat hidup, bertahan hidup, dan memperbanyak diri di dalam sel inang. Metode pemberian vaksin DNA secara oral melalui <em>Artemia</em> sp. merupakan salah satu alternatif pengobatan yang diharapkan dapat menangani permasalahan penyakit pada ikan yang disebabkan oleh virus. Pada penelitian ini dilakukan uji ekspresi vaksin DNA yang menyandikan glikoprotein 11 (GP-11) pada ikan mas. Bakteri yang mengandung plasmid Krt-GP-11 sebagai vaksin diberikan melalui <em>Artemia</em> sp. sebagai pembawa vaksin. Pemberian <em>Artemia</em> sp. dilakukan satu dan dua kali seminggu pada ikan mas umur tiga minggu. Keberadaan DNA vaksin di usus, ginjal, dan insang dianalisis menggunakan metode PCR. Organ diambil setiap tiga hari setelah pemberian vaksin. Ekspresi gen GP-11 juga diamati pada organ ginjal di setiap perlakuan dengan menggunakan metode RT-PCR, pada sepuluh hari setelah pemberian vaksin. Hasil penelitian menunjukkan bahwa DNA vaksin yang diberikan dengan dosis 10<sup>6</sup> cfu/mL pada perlakuan satu dan dua kali seminggu dapat terdeteksi pada ketiga organ. Hasil RT-PCR menunjukkan bahwa ekspresi GP-11 dapat terdeteksi pada semua organ uji di setiap perlakuan. Dengan demikian <em>Artemia</em> sp. dapat digunakan sebagai vektor pembawa vaksin plasmid GP-11 dengan frekuensi pemberian vaksin untuk larva ikan mas.</p><p class="NoParagraphStyle"> </p><p>Kata kunci: vaksin DNA, KHV, <em>Artemia</em> sp., ikan mas</p>

scholarly journals Rapid non- invasive detection of Influenza- A- infection by multicapillary column coupled ion mobility spectrometry

2020 ◽  
Author(s):  
Claus Steppert ◽  
Isabel Steppert ◽  
Thomas Bollinger ◽  
William Sterlacci
Keyword(s):  
Ion Mobility Spectrometry ◽  
Ion Mobility ◽  
Influenza A ◽  
Viral Infections ◽  
Multivariate Statistical ◽  
Infected People ◽  
Staff Members ◽  
Number Of Patients ◽  
Registration Number ◽  
Multicapillary Column

Infectious pathogens are a global issue. Global air travelling offers an easy and fast opportunity not only for people but also for infectious diseases to spread around the world within a few days. Also, large public events facilitate increasing infection numbers. Therefore, a rapid on-site screening for infected people is urgently needed. Due to the small size and easy handling, the ion mobility spectrometry coupled with a multicapillary column (MCC-IMS) is a very promising, sensitive method for the on-site identification of infectious pathogens based on scents, representing volatile organic compounds (VOCs). The purpose of this study was to prospectively assess whether identification of Influenza-A- infection based on VOCs by MCC-IMS is possible in breath. Nasal breath was investigated in 24 consecutive persons with and without Influenza-A-infection by MCC-IMS. In 14 Influenza-A-infected patients, infection was proven by PCR of nasopharyngeal swabs. Four healthy staff members and six patients with negative PCR result served as controls. For picking up relevant VOCs in MCC-IMS spectra, software based on cluster analysis followed by multivariate statistical analysis was applied. With only four VOCs canonical discriminant analysis was able to distinguish Influenza-A-infected patients from not infected with 100% sensitivity and 100% specificity. This present proof-of-concept- study yields encouraging results showing a rapid diagnosis of viral infections in nasal breath within 5 minutes by MCC-IMS. The next step is to validate the results with a greater number of patients with Influenza-A-infection as well as other viral diseases, especially COVID-19. Registration number at ClinicalTrials.gov NCT04282135.

scholarly journals Molecular detection of Mycobacterium tuberculosis in pulmonary and extrapulmonary samples in a hospital-based study

2020 ◽  
Vol 20 (4) ◽  
pp. 1617-23
Author(s):  
Kalal Iravathy Goud ◽  
Matam Kavitha ◽  
Adi Mahalakshmi ◽  
Ravi Vempati ◽  
Abdulaziz A Alodhayani ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pcr Analysis ◽  
Sequencing Analysis ◽  
Rt Pcr ◽  
Tissue Samples ◽  
Non Invasive ◽  
Invasive Test ◽  
Keywords Tuberculosis ◽  
Bronchoalveolar Fluid

Objective: Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a deadly infectious disease. India contributes to one-third of the global TB burden. However, no studies have been carried out in the Telangana (Hyderabad) population using real-time polymerase chain reaction (RT-PCR). Therefore, the current study evaluated the role of RT-PCR as a rapid and non-invasive test to diagnose TB by testing for pulmonary tuberculosis (PTB) and extrapulmonary tubercu- losis (EPTB). Materials and methods: This hospital-based study examined 1670 samples (900 EPTB; 770 PTB) comprising tissue (n = 537), peritoneal fluid (n = 420), sputum (n = 166), bronchial fluid (n = 126), cerebrospinal fluid (n = 145), ascetic fluid (n = 76), sputum pus (n = 78), urine (n = 79), and bronchoalveolar fluid (n = 43) samples. DNA from samples was separated using specific isolation kits and subjected to RT-PCR. Results: In this study, we enrolled 1670 subjects and categorized 54.4% as females and 45.6% as males. The collected sam- ples were categorized as 48.5% of fluid samples, followed by tissue (32.2%), sputum (9.9%), urine (4.7%), and pus-swab (4.6%). RT-PCR analysis revealed that 4.7% patients were positive for Mtb. Our results revealed that 61% of the affected patients were male and 39% were female. Among the specimen types, tissue samples gave the highest proportion of positive results (36.3%). Conclusion: The results showed that RT-PCR should be implemented and given top priority in TB diagnosis to save time and facilitate a definitive diagnosis. Tissue samples are highly recommended to screen the Mtb through the technique RT- PCR. Future studies should extend the technique to the global population and exome sequencing analysis should be per- formed to identify TB risk markers. Keywords: Tuberculosis (TB); EPTB; PTB; Mycobacterium tuberculosis (Mtb).

scholarly journals Does sampling saliva increase detection of SARS-CoV-2 by RT-PCR? Comparing saliva with oro-nasopharyngeal swabs

2020 ◽  
Author(s):  
Ozlem Akgun Dogan ◽  
Betsi Kose ◽  
Nihat Bugra Agaoglu ◽  
Jale Yildiz ◽  
Gizem Alkurt ◽  
...  
Keyword(s):  
Gold Standard ◽  
Viral Rna ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Standard Samples ◽  
Rna Detection ◽  
Detection Rates ◽  
Gold Standard Method ◽  
Non Invasive ◽  
Isolation Period

The gold standard method in the diagnosis of SARS-CoV-2 infection is the detection of viral RNA in nasopharyngeal sample by RT-PCR. Recently, saliva samples has been suggested as an alternative due to being fast, reliable and non-invasive, rather than nasopharyngeal samples. We compared RT-PCR results in nasopharyngeal, oro-nasopharyngeal and saliva samples of COVID-19 patients. 98 of 200 patients were positive in RT-PCR analysis performed before the hospitalization. In day 0, at least one sample was positive in 67% of 98 patients. Positivity rate was 83% for both oro-nasopharyngeal and nasopharyngeal samples, while it was 63% for saliva samples (p<0.001). On day 5, RT-PCR was performed in 59 patients, 34% had at least one positive result. The positivity rate was 55% for saliva and nasopharyngeal samples, while it was 60% for oro-nasopharyngeal samples. Our study shows that the sampling saliva does not increase the sensitivity of RT-PCR tests at early stages of infection. However, on 5th day, viral RNA detection rates in saliva were similar to nasopharyngeal and oro-nasopharyngeal samples. In conclusion, we suggest that, in patients receiving treatment, virus presence in saliva, in addition to the standard samples, is important to determine the isolation period and to control the transmission.

Discovery of molecular biomarker signatures for the detection of bladder cancer.

2013 ◽  
Vol 31 (6_suppl) ◽  
pp. 302-302
Author(s):  
Evan Gomes ◽  
Virginia Urquidi ◽  
Yunpeng Cai ◽  
Yijun Sun ◽  
Charles Joel Rosser ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Disease Status ◽  
Healthcare Systems ◽  
Urothelial Cell ◽  
Pcr Analysis ◽  
Monitoring And Control ◽  
Rt Pcr ◽  
Non Invasive ◽  
Diagnostic Signature ◽  
Independent Cohort

302 Background: Bladder cancer (BCa) is among the five most common malignancies world-wide, and due to high rates of recurrence, one of the most prevalent. Improvements in non-invasive urine-based assays to detect BCa would benefit both patients and healthcare systems. In this study, the goal was to identify urothelial cell transcriptomic signatures associated with BCa. Methods: Gene expression profiling (Affymetrix U133 Plus 2.0 arrays) was applied to exfoliated urothelia obtained from a cohort of 92 subjects with known bladder disease status. Computational analyses identified candidate biomarkers of BCa and an optimal predictive model was derived. Selected targets from the profiling analyses were monitored in an independent cohort of 81 subjects using quantitative real-time PCR (RT-PCR). Results: Data analysis identified 52 genes associated with BCa (p≤0.001), and gene models that optimally predicted class label were derived. RT-PCR analysis of 48 selected targets in an independent cohort identified a 14-gene diagnostic signature that predicted the presence of BCa with a specificity of 100% at 90% sensitivity. Conclusions: Exfoliated urothelia sampling provides a robust analyte for the evaluation of patients with suspected BCa. The refinement and validation of the multi-gene urothelial cell signatures identified in this preliminary study may lead to accurate, non-invasive assays for the detection of BCa. The development of an accurate, non-invasive BCa detection assay would benefit both the patient and healthcare systems through better detection, monitoring and control of disease.

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