Q fever: A neglected disease of camels in Giza and Cairo Provinces, Egypt

Background and Aim: Q fever is a zoonotic disease caused by Coxiella burnetii. Cattle, sheep, and goat are the main reservoir of C. burnetii. In Egypt, the epidemiological data about C. burnetii in camels are limited. Therefore, the current study was conducted to identify C. burnetii infection in camels by different molecular tools and to estimate its seropositivity through the detection of anti-C. burnetii antibodies in camel sera. Materials and Methods: Blood samples were collected 112 from camels in Giza and Cairo Provinces, Egypt. All blood samples were screened by trans-quantitative polymerase chain reaction (trans-qPCR) for C. burnetii and positive samples subjected to standard PCR using the superoxide dismutase enzyme coding gene of C. burnetii. Sera of studied camels were examined for the presence of antibodies against C. burnetii using enzyme-linked immunosorbent assay. Results: Out of 112 camels, 19 were positive for C. burnetii by qPCR with an overall prevalence of 16.9% (18.6% in Giza and 15.1% in Cairo Provinces, respectively). The seroprevalence of anti-C. burnetii IgG antibodies in the examined camels was 4.5% (5/112). Conclusion: Trans-qPCR assay is a rapid and sensitive tool for the detection of C. burnetii in acute stage. Camels should be considered one of the major reservoirs for C. burnetii in Egypt.

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Diagnostic accuracy of Fusobacterium nucleatum IgA and IgG ELISA test in colorectal cancer

Scientific Reports ◽

10.1038/s41598-021-81171-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Melike Kurt ◽

Zeki Yumuk

Keyword(s):

Colorectal Cancer ◽

Quantitative Polymerase Chain Reaction ◽

Invasive Procedure ◽

Fusobacterium Nucleatum ◽

Elisa Test ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Discriminative Ability ◽

Non Invasive ◽

Polymerase Chain

AbstractThe colorectal cancer is a serious health problem. The diagnosis of the disease mostly relies on an invasive procedure. A non-invasive diagnostic test such as an immunoassay, may facilitate diagnosis of colorectal cancer. The purpose of the study was to evaluate the use of antibodies against Fusobacterium nucleatum in the diagnosis of colorectal cancer (CRC). Totally 78 patients in three groups were included in the study. F. nucleatum in the tissues was detected using quantitative polymerase chain reaction assay. F. nucleatum IgA and IgG were measured using enzyme linked immunosorbent assay. F. nucleatum was detected in 86.7% and 73.1% cases of CRC and precancerous-benign colon disease (P-BCD), respectively. The OD values from F. nucleatum IgA and IgG ELISA tests were higher in CRC group compared with healthy individuals. The sensitivity of IgA ELISA test varied between 31.8 and 95.5% depending on the chosen cut-off values. The positivity rate of antibodies in patients with high amount of F. nucleatum in tissue was significantly greater than in the negative group. The F. nucleatum IgA and IgG antibodies in CRC were higher than the ones in healthy controls but the discriminative ability of the ELISA test was not adequate to be considered as a diagnostic tool.

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Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2408 ◽

2021 ◽

Vol 11 (3) ◽

pp. 373-379

Author(s):

Huitao Li ◽

Xueyu Chen ◽

Xiaomei Qiu ◽

Weimin Huang ◽

Chuanzhong Yang

Keyword(s):

Polymerase Chain Reaction ◽

Limit Of Detection ◽

Quantitative Polymerase Chain Reaction ◽

Diagnostic Methods ◽

Chain Reaction ◽

Fungal Isolation ◽

Blood Samples ◽

Fungal Strains ◽

Polymerase Chain ◽

Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

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Measles antibodies and response to vaccination in children aged less than 14 months: implications for age of vaccination

Epidemiology and Infection ◽

10.1017/s0950268811002184 ◽

2011 ◽

Vol 140 (9) ◽

pp. 1599-1606 ◽

Cited By ~ 16

Author(s):

E. BORRÀS ◽

L. URBIZTONDO ◽

J. COSTA ◽

J. BATALLA ◽

N. TORNER ◽

...

Keyword(s):

Immunosorbent Assay ◽

Maternal Antibodies ◽

Passive Immunity ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Blood Samples ◽

Measles Outbreak ◽

Measles Antibodies ◽

Measles Vaccination

SUMMARYPassive immunity against measles decreases during the first months of life. The objective of this study was to determine titres of measles antibodies in children aged 9–14 months and their mothers before vaccination, and the children's response to vaccination. Blood samples were collected by capillary puncture before and 28 days after vaccination. Samples were obtained between February and June 2007 during an ongoing measles outbreak. Titres of specific measles IgG antibodies were determined by enzyme-linked immunosorbent assay. Seroconversion was defined as the presence of antibodies after vaccination in subjects without antibodies before vaccination. Maternal antibodies were present in 37·7% of all 69 children included and in 45·1% of children aged 9 months. Of the 51 children in whom a second sample was obtained, 31 (60·8%) were seronegative before vaccination and 61·3% seroconverted. Interference of maternal antibodies was 30%. Advancing the first dose of measles vaccination from 15 to 12 months is a correct strategy, given the increase in the time of susceptibility of infants to measles.

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Prevalence and Molecular Epidemiological Data onDirofilaria immitisin Dogs from Northeastern States of India

The Scientific World JOURNAL ◽

10.1155/2015/265385 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 10

Author(s):

Sonjoy Kumar Borthakur ◽

Dilip Kumar Deka ◽

Saidul Islam ◽

Dilip Kumar Sarma ◽

Prabhat Chandra Sarmah

Keyword(s):

Epidemiological Data ◽

Elisa Test ◽

Molecular Techniques ◽

Diagnostic Methods ◽

Molecular Tools ◽

Stray Dogs ◽

Concentration Technique ◽

Working Dogs ◽

Polymerase Chain

The aim of the present study was to determine the prevalence ofDirofilaria immitisin stray, pet, and working dogs (n=413, 266, and 103, resp.) from Guwahati (Assam) and Aizawl (Mizoram), areas located in two Northeastern States of India. Diagnostic methods applied were microscopy (wet film and Knott’s concentration technique), immunological test (Ag ELISA by SNAP 4Dx ELISA kit), and molecular tools (polymerase chain reaction and sequencing), which evidenced 11.38, 18.03, and 13.93% of positive animals, respectively. No significant differences were observed by area (18.23% versus 17.68%) nor by sex (18.1% versus 17.9%), whereas stray dogs proved more infected than other groups (P<0.05). ELISA test evidenced an overall 22.69% of occult infections, mainly in working dogs (60%), and molecular techniques detectedDirofilaria (Nochtiella) repensin 4 stray dogs from Guwahati. Characterization ofD. immitisisolates for ITS-2 region showed close identity with South Asian isolates.

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Further studies of neuroangiostrongyliasis (rat lungworm disease) in Australian dogs: 92 new cases (2010–2020) and results for a novel, highly sensitive qPCR assay

Parasitology ◽

10.1017/s0031182020001572 ◽

2020 ◽

pp. 1-9

Author(s):

Rogan Lee ◽

Tsung-Yu Pai ◽

Richard Churcher ◽

Sarah Davies ◽

Jody Braddock ◽

...

Keyword(s):

Infectious Disease ◽

Quantitative Polymerase Chain Reaction ◽

Enzyme Linked Immunosorbent Assay ◽

Chain Reaction ◽

Cell Counts ◽

Highly Sensitive ◽

Elisa Testing ◽

Rat Lungworm ◽

Polymerase Chain ◽

Sydney Australia

Abstract The principal aim of this study was to optimize the diagnosis of canine neuroangiostrongyliasis (NA). In total, 92 cases were seen between 2010 and 2020. Dogs were aged from 7 weeks to 14 years (median 5 months), with 73/90 (81%) less than 6 months and 1.7 times as many males as females. The disease became more common over the study period. Most cases (86%) were seen between March and July. Cerebrospinal fluid (CSF) was obtained from the cisterna magna in 77 dogs, the lumbar cistern in f5, and both sites in 3. Nucleated cell counts for 84 specimens ranged from 1 to 146 150 cells μL−1 (median 4500). Percentage eosinophils varied from 0 to 98% (median 83%). When both cisternal and lumbar CSF were collected, inflammation was more severe caudally. Seventy-three CSF specimens were subjected to enzyme-linked immunosorbent assay (ELISA) testing for antibodies against A. cantonensis; 61 (84%) tested positive, titres ranging from <100 to ⩾12 800 (median 1600). Sixty-one CSF specimens were subjected to real-time quantitative polymerase chain reaction (qPCR) testing using a new protocol targeting a bioinformatically-informed repetitive genetic target; 53/61 samples (87%) tested positive, C T values ranging from 23.4 to 39.5 (median 30.0). For 57 dogs, it was possible to compare CSF ELISA serology and qPCR. ELISA and qPCR were both positive in 40 dogs, in 5 dogs the ELISA was positive while the qPCR was negative, in 9 dogs the qPCR was positive but the ELISA was negative, while in 3 dogs both the ELISA and qPCR were negative. NA is an emerging infectious disease of dogs in Sydney, Australia.

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Molecular, serological and epidemiological observations after a suspected outbreak of plague in Nyimba, eastern Zambia

Tropical Doctor ◽

10.1177/0049475516662804 ◽

2016 ◽

Vol 47 (1) ◽

pp. 38-43 ◽

Cited By ~ 4

Author(s):

Stanley S Nyirenda ◽

Bernard M Hang’ombe ◽

Bukheti S Kilonzo ◽

Mathews N Kabeta ◽

Mundia Cornellius ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Yersinia Pestis ◽

Plasminogen Activator ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Recent Past ◽

Chain Reaction ◽

Polymerase Chain ◽

Pcr Techniques ◽

Suspected Outbreak

Plague is a re-emerging zoonotic disease caused by the bacterium Yersinia pestis. The disease has caused periodic global devastation since the first outbreak in the 6th century. Two months after a suspected plague outbreak in Nyimba district, samples were collected from 94 livestock (goats and pigs), 25 rodents, 6 shrews and 33 fleas. Enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) techniques were used to investigate the presence of Y. pestis, which showed that 16.0% (4/25) of rodents, 16.7% (1/6) of shrews ( Crocidura spp) and 6.0% (5/83) of goats were positive for IgG antibodies against Fraction 1 antigen of Y. pestis. Plasminogen activator (Pla) gene (DNA) of Y. pestis was detected in five pools containing 36.4% (12/33) fleas collected from pigs (n = 4), goats (n = 5) and rodents (n = 3). The detection of Pla gene in fleas and IgG antibodies against Fraction1 antigen in rodents, shrews and goats suggest that Y. pestis had been present in the study area in the recent past.

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Comparison of culture and qPCR methods in detection of mycobacteria from drinking waters

Canadian Journal of Microbiology ◽

10.1139/cjm-2012-0695 ◽

2013 ◽

Vol 59 (4) ◽

pp. 280-286 ◽

Cited By ~ 5

Author(s):

Noora H.J. Räsänen ◽

Helena Rintala ◽

Ilkka T. Miettinen ◽

Eila Torvinen

Keyword(s):

Surface Water ◽

Quantitative Polymerase Chain Reaction ◽

Water Source ◽

Assimilable Organic Carbon ◽

Environmental Mycobacteria ◽

Drinking Waters ◽

Qpcr Method ◽

Sensitive Tool ◽

Different Types ◽

Polymerase Chain

Environmental mycobacteria are common bacteria in man-made water systems and may cause infections and hypersensitivity pneumonitis via exposure to water. We compared a generally used cultivation method and a quantitative polymerase chain reaction (qPCR) method to detect mycobacteria in 3 types of drinking waters: surface water, ozone-treated surface water, and groundwater. There was a correlation between the numbers of mycobacteria obtained by cultivation and qPCR methods, but the ratio of the counts obtained by the 2 methods varied among the types of water. The qPCR counts in the drinking waters produced from surface or groundwater were 5 to 34 times higher than culturable counts. In ozone-treated surface waters, both methods gave similar counts. The ozone-treated drinking waters had the highest concentration of assimilable organic carbon, which may explain the good culturability. In warm tap waters, qPCR gave 43 times higher counts than cultivation, but both qPCR counts and culturable counts were lower than those in the drinking waters collected from the same sites. The TaqMan qPCR method is a rapid and sensitive tool for total quantitation of mycobacteria in different types of clean waters. The raw water source and treatments affect both culturability and total numbers of mycobacteria in drinking waters.

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APicrorhiza kurroaDerivative, Picroliv, Attenuates the Development of Dextran-Sulfate-Sodium-Induced Colitis in Mice

Mediators of Inflammation ◽

10.1155/2012/751629 ◽

2012 ◽

Vol 2012 ◽

pp. 1-9 ◽

Cited By ~ 9

Author(s):

De-Kui Zhang ◽

Jian-Jie Yu ◽

Yu-Min Li ◽

Li-Na Wei ◽

Yi Yu ◽

...

Keyword(s):

Dextran Sulfate ◽

Dextran Sulfate Sodium ◽

Activity Index ◽

Quantitative Polymerase Chain Reaction ◽

Critical Role ◽

Disease Activity Index ◽

Enzyme Linked Immunosorbent Assay ◽

Histological Score ◽

Polymerase Chain ◽

Histology Score

Background. Free radicals and proinflammatory cytokines have been shown to play a critical role in the pathogenesis of ulcerative colitis (UC). Picroliv, aPicrorhiza kurroaderivative, has been demonstrated to have antioxidant and anti-inflammatory effect. The purpose of the study was to investigate the effects of picroliv on experimental model of UC in mice.Materials and Methods. Picroliv was administrated orally by gavage to mice with colitis induced by dextran sulfate sodium (DSS). Disease activity index (DAI), colon length, and histology score were observed. Myeloperoxidase (MPO) activity, and SOD, MDA concentrations were measured by enzyme-linked immunosorbent assay (ELISA) while the expression of cytokine mRNAs was studied by real-time-quantitative polymerase chain reaction and also ELISA. The expression of NF-κB p65 was observed by immunohistochemistry staining and western blotting.Results. A significant improvement was observed in DAI and histological score in mice treated with picroliv, and incerased MPO activity, MDA concentrations, and the expression of IL-1β, TNF-α, and NF-κB p65 in mice with DSS-induced colitis were significantly reduced while decreased SOD level increased following administration of picroliv.Conclusion. The administration of picroliv leads to an amelioration of DSS-induced colitis, suggesting administration of picroliv may provide a therapeutic approach for UC.

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Biodistribution and serologic response in SARS-CoV-2 induced ARDS: A cohort study

PLoS ONE ◽

10.1371/journal.pone.0242917 ◽

2020 ◽

Vol 15 (11) ◽

pp. e0242917

Author(s):

Tobias Schlesinger ◽

Benedikt Weißbrich ◽

Florian Wedekink ◽

Quirin Notz ◽

Johannes Herrmann ◽

...

Keyword(s):

Intensive Care ◽

Viral Load ◽

Disease Severity ◽

Quantitative Polymerase Chain Reaction ◽

Antiviral Treatment ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Intensive Care Treatment ◽

Blood Samples ◽

Tracheal Aspirates

Background The viral load and tissue distribution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remain important questions. The current study investigated SARS-CoV-2 viral load, biodistribution and anti-SARS-CoV-2 antibody formation in patients suffering from severe corona virus disease 2019 (COVID-19) induced acute respiratory distress syndrome (ARDS). Methods This is a retrospective single-center study in 23 patients with COVID-19-induced ARDS. Data were collected within routine intensive care. SARS-CoV-2 viral load was assessed via reverse transcription quantitative polymerase chain reaction (RT-qPCR). Overall, 478 virology samples were taken. Anti-SARS-CoV-2-Spike-receptor binding domain (RBD) antibody detection of blood samples was performed with an enzyme-linked immunosorbent assay. Results Most patients (91%) suffered from severe ARDS during ICU treatment with a 30-day mortality of 30%. None of the patients received antiviral treatment. Tracheal aspirates tested positive for SARS-CoV-2 in 100% of the cases, oropharyngeal swabs only in 77%. Blood samples were positive in 26% of the patients. No difference of viral load was found in tracheal or blood samples with regard to 30-day survival or disease severity. SARS-CoV-2 was never found in dialysate. Serologic testing revealed significantly lower concentrations of SARS-CoV-2 neutralizing IgM and IgA antibodies in survivors compared to non-survivors (p = 0.009). Conclusions COVID-19 induced ARDS is accompanied by a high viral load of SARS-CoV-2 in tracheal aspirates, which remained detectable in the majority throughout intensive care treatment. Remarkably, SARS-CoV-2 RNA was never detected in dialysate even in patients with RNAemia. Viral load or the buildup of neutralizing antibodies was not associated with 30-day survival or disease severity.

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Evaluation of anti-SARS-CoV-2 antibody testing in asymptomatic or mild COVID-19 patients in outbreak on a cruise ship

10.1101/2021.03.10.21253064 ◽

2021 ◽

Author(s):

Norihito Kaku ◽

Fumitaka Nishimura ◽

Yui Shigeishi ◽

Rina Tachiki ◽

Hironori Sakai ◽

...

Keyword(s):

Genetic Test ◽

False Negative ◽

Enzyme Linked Immunosorbent Assay ◽

Cruise Ship ◽

Igg Antibodies ◽

Antibody Testing ◽

Predictive Values ◽

Blood Samples ◽

Negative Results ◽

Genetic Test Results

AbstractBackgroundA few studies on antibody testing have focused on asymptomatic or mild coronavirus disease 2019 (COVID-19) patients with low initial anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody responses. Anti-SARS-CoV-2 antibody-testing performance was evaluated using blood samples from asymptomatic or mild COVID-19 patients.MethodsBlood samples were collected from 143 COVID-19 patients during an outbreak on a cruise ship 3 weeks after diagnosis. Simultaneously, a second SARS-CoV-2 genetic test was performed. Samples stored before the COVID-19 pandemic were also used to evaluate the lateral flow immunochromatographic assay (LFA) and electrochemiluminescence immunoassay (ECLIA). Titers of anti-SARS-CoV-2 IgM and IgG antibodies against the nucleocapsid and spike proteins were measured using the enzyme-linked immunosorbent assay to compare false-negative- with positive-result samples.ResultsSensitivity, specificity, positive-predictive, and negative-predictive values of LFA-detected IgM antibodies were 0.231, 1.000, 1.000, and 0.613, respectively; those of LFA-detected IgG antibodies were 0.483, 0.989, 0.972, and 0.601, respectively; and those of ECLIA-detected total antibodies were 0.783, 1.000, 1.000, and 0.848, respectively. IgM-, IgG-, and total-antibody positivity rates in the patients with negative results from the second genetic testing were 22.9%, 47.6%, and 72.4%, respectively. All antibody titers, especially those of the IgG antibody against nucleocapsid protein, were significantly lower in blood samples with false-negative results than in those with positive results.ConclusionsThese findings suggest that anti-SARS-CoV-2 antibody testing has lower performance in asymptomatic or mild COVID-19 patients than required in the guidelines, and situations in which it is useful are limited.Key pointsAnti-SARS-CoV-2 antibody testing in asymptomatic or mild COVID-19 patients is lower than the required clinical sensitivity, although it may be useful in patients at 3–4 weeks after symptom onset but with negative SARS-CoV-2 genetic test results.

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