False-negative reactions to the comparative intradermal tuberculin test for bovine tuberculosis

ABSTRACT: According to the Brazilian National Program for the Control and Eradication of Animal Brucellosis and Tuberculosis (PNCEBT), the routine tests for the diagnosis of bovine tuberculosis in the country are the simple intradermal tuberculin test (SITT) of the Ministry of Agriculture, Livestock and Food Supply (MAPA), the caudal fold test and the comparative intradermal tuberculin test (CITT). The latter is also used as a confirmatory test. A group of 53 animals from three dairy herds in a focal area for bovine tuberculosis, that were submitted to depopulation in the state of Rio Grande do Sul, were submitted to the CITT. Tissues were cultured and the resulting colonies were confirmed by PCR and DNA sequencing. Among the 53 animals analyzed using the CITT, 32 (60.4%) were negative, 14 (26.4%) were positive and seven (13.2%) results were inconclusive. The CITT detected 11 of the 39 animals with culture-confirmed M. bovis infection as positive. Among the total of 14 uninfected animals based on cultures, the CBT detected eight as negative. Thus, the CITT demonstrated sensitivity of 28.2% and specificity of 57.1% for the population sampled. A total of 24/32 (75.0%) of the animals with negative CITT results were culture positive (confirmed by PCR) and were considered false negatives based on the CITT. The maintenance of these false-negative animals in herds has serious implications for the control of the disease, since they can be a source of infection. The addition of complementary tests could help identify such animals and increase the odds of diagnostic success.

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Host Serum Proteins as Potential Biomarkers of Bovine Tuberculosis Resistance Phenotype

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.734087 ◽

2021 ◽

Vol 8 ◽

Author(s):

Jorge Luis Mazorra-Carrillo ◽

Omar Antonio Alcaraz-López ◽

Gonzalo López-Rincón ◽

Bernardo Villarreal-Ramos ◽

José A. Gutiérrez-Pabello ◽

...

Keyword(s):

Bovine Tuberculosis ◽

Serum Proteins ◽

Amine Oxidase ◽

Tuberculin Test ◽

Ifn Gamma ◽

Holstein Friesian ◽

Novel Biomarkers ◽

Primary Amine Oxidase ◽

Friesian Cows ◽

Control And Eradication

Eradication of bovine tuberculosis (bTB) continues to be a worldwide challenge. The lack of reliable vaccines dampens the control and eradication programs of Mycobacterium bovis infection and spread. Selection and breeding of cattle resistant to M. bovis infection would greatly enhance the effectiveness of bTB eradication programs. Here, we have evaluated the potential of serum proteins as biomarkers of cattle resistance to bTB in Holstein-Friesian cows, 6–8-year-old, born and raised in similar conditions in herds with bTB prevalence >30%. Serum proteins obtained from uninfected cows (bTB-resistant; R) were compared to those from infected cows (bTB-susceptible; S), defined by a negative or positive bTB diagnosis, respectively. bTB diagnosis included: (i) single intradermal (caudal fold) tuberculin test, (ii) whole blood IFN-gamma test, (iii) gross visible lesions in lymph nodes and lungs by inspection at the abattoir, and (iv) a bacteriological culture for M. bovis. Using 2D-GE and LC-ESI-MS/MS, we found higher expression levels of primary amine oxidase (AO), complement component 5 (C5), and serotransferrin (TF) in R cattle than S cattle. In-house developed and standardized ELISAs for these novel biomarkers showed the best sensitivities of 72, 77, 77%, and specificities of 94, 94, 83%, for AO, C5, and TF, respectively. AUC-ROC (95% CI) values of 0.8935 (0.7906–0.9964), 0.9290 (0.8484–1.010), and 0.8580 (0.7291–0.9869) were obtained at cut-off points of 192.0, 176.5 ng/ml, and 2.1 mg/ml for AO, C5, and TF, respectively. These proteins are involved in inflammatory/immunomodulatory responses to infections and may provide a novel avenue of research to determine the mechanisms of protection against bTB. Overall, our results indicate that these proteins could be novel biomarkers to help identify cattle resistant to bTB, which in turn could be used to strengthen the effectiveness of existing eradication programs against bTB.

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Bovine tuberculosis in domestic and wild mammals in an area of Dorset. I. Tuberculosis in cattle

Journal of Hygiene ◽

10.1017/s002217240007073x ◽

1982 ◽

Vol 89 (2) ◽

pp. 195-210 ◽

Cited By ~ 34

Author(s):

J. W. Wilesmith ◽

T. W. A. Little ◽

H. V. Thompson ◽

C. Swan

Keyword(s):

Mycobacterium Bovis ◽

Bovine Tuberculosis ◽

Tuberculin Test ◽

Common Source ◽

Wild Mammals ◽

Source Of Infection ◽

Short Period ◽

Major Outbreak ◽

Two Peaks

SUMMARYA major outbreak of tuberculosis occurred in cattle on a farm in Dorset between 1970 and 1976. Six hundred and twenty-six cattle were slaughtered either because they reacted to the tuberculin test or had been exposed to infection. No source of infection was found until 1974 when badgers infected with Mycobacterium bovis were first discovered.An analysis of the tuberculin test records of this herd and the six surrounding herds indicated that tuberculosis had been a sporadic problem since the early 1960's. Two peaks of infection occurred in the most severely affected herd in 1970 and 1974 when 29·8% and 27·3% of animals, respectively, reacted to the tuberculin test. These figures are exceptionally high. During the last 20 years there have been two periods when all the herds in the area had synchronous outbreaks consistent with a common source.Analysis indicated that cattle were at greatest risk in April and May and suggest that there was re-exposure to infection at this time each year. In addition the cattle were apparently exposed to M. bovis, at sufficiently high levels for transmission to occur, for only a relatively short period of time.

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Epidemiological status of bovine tuberculosis in the State of Rio Grande do Sul, Brazil

Semina Ciências Agrárias ◽

10.5433/1679-0359.2016v37n5supl2p3647 ◽

2016 ◽

Vol 37 (5Supl2) ◽

pp. 3647 ◽

Cited By ~ 2

Author(s):

Mariana Ramos Queiroz ◽

Ana Cláudia Mello Groff ◽

Nariléia Dos Santos Silva ◽

José Henrique Hildebrand Grisi-Filho ◽

Marcos Amaku ◽

...

Keyword(s):

Risk Factors ◽

Bovine Tuberculosis ◽

Dairy Herd ◽

Rio Grande ◽

Tuberculin Test ◽

The State ◽

Surveillance Systems ◽

Rio Grande Do Sul ◽

Factors Associated ◽

Minimum Interval

A study was conducted to determine the epidemiological status of bovine tuberculosis in the state of Rio Grande do Sul. The state was divided in seven regions, and in each of them, a pre-established number of farms was randomly sampled. In each farm, cows with age equal to or greater than 24 months were selected at random and submitted to the comparative cervical tuberculin test. The animals whose tests were inconclusive were retested with the same diagnostic procedure within a minimum interval of 60 days. In all, 9,895 animals from 1,067 farms were tested. An epidemiological questionnaire was applied in the farms in order to identify risk factors associated with bovine tuberculosis. The prevalence of infected herds in the state was 2.8% [1.8; 4.0] and that of infected animals was 0.7% [0.4; 1.0]. There was a trend towards a concentration of infected herds in the northern part of the state, with a predominance of dairy and mixed herds. The risk factors associated with the condition of infected herds were being a dairy herd (OR = 2.90 [1.40; 6.13]) and herds with 16 or more cows (OR = 2.61 [1.20; 5.49]). Thus, the best strategy to be adopted by the state is the implementation of surveillance systems to detect and remediate the infected herds, preferably incorporating elements of risk-based surveillance. In addition, the state must carry out a solid action of health education so that the producers test animals for bovine tuberculosis before introducing them in their herds.

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Study on supplemental test to improve the detection of bovine tuberculosis in individual animals and herds

BMC Veterinary Research ◽

10.1186/s12917-021-02839-4 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Paulo Alex Machado Carneiro ◽

Eliomar de Moura Sousa ◽

Rinaldo Batista Viana ◽

Bruno Moura Monteiro ◽

Aline do Socorro Lima Kzam ◽

...

Keyword(s):

Mycobacterium Bovis ◽

Bovine Tuberculosis ◽

False Negative ◽

Tuberculin Test ◽

Enzyme Linked Immunosorbent Assay ◽

Late Nineteenth Century ◽

Routine Testing ◽

Control Programs ◽

Increased Sensitivity ◽

And Control

Abstract Background Bovine tuberculosis (bTB), is a worldwide disease caused by Mycobacterium bovis (M. bovis). The success of bTB eradication and control programs is based on early detection and the removal of reactors from a herd thus routine testing and cull strategy have been applied globally. Since the late nineteenth century, the Tuberculin Skin Test (TST) has been the primary antemortem test available to support bTB eradication campaigns. Due to the TST limitations in terms of Se and Sp, the credibility of the diagnosis is frequently questioned given the occurrence of false-positive and false-negative reactions, therefore, it is necessary to confirm reactive animals using other methods, ensuring the reliability of the diagnosis. The aim of this study was to evaluate the sensitivity and specificity of a multiplex enzyme-linked immunosorbent assay (ELISA) relative to the tuberculin test used for the diagnosis of tuberculosis in cattle in Brazil. Results Lack of agreement between comparative cervical tuberculin test and ELISA IDEXX TM was observed. The 2 animals positive on the comparative cervical tuberculin test did not react at the ELISA IDEXX TM and 22 negative reactors by comparative cervical tuberculin test were positive by the ELISA IDEXX TM. The ELISA IDEXX TM showed sensitivity that is significantly lower than the official screening test the single cervical tuberculin. ELISA IDEXX TM also detected infected animals and herds undetected by the comparative cervical tuberculin test. The parallel use of comparative cervical tuberculin test and ELISA IDEXX TM increased sensitivity and the feasibility bTB screening. Conclusions The results obtained here suggest that the ELISA IDEXX TM may be a supplemental test for the detection of Mycobacterium bovis infection in regions without routine testing and slaughter, where the disease generally progresses to more advanced stages and antibody responses are likely to be more prevalent. Evidence to support the validation of the ELISA IDEXX™ as a supplemental test for bTB eradication programs was provided.

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Clinically Meaningful Change

Methodology ◽

10.1027/1614-2241/a000168 ◽

2019 ◽

Vol 15 (3) ◽

pp. 97-105

Author(s):

Rodrigo Ferrer ◽

Antonio Pardo

Keyword(s):

Effect Size ◽

False Negative ◽

False Negative Rate ◽

Point Of View ◽

Skewed Distribution ◽

Effect Sizes ◽

False Negatives ◽

Large Size ◽

Before And After ◽

Post Test

Abstract. In a recent paper, Ferrer and Pardo (2014) tested several distribution-based methods designed to assess when test scores obtained before and after an intervention reflect a statistically reliable change. However, we still do not know how these methods perform from the point of view of false negatives. For this purpose, we have simulated change scenarios (different effect sizes in a pre-post-test design) with distributions of different shapes and with different sample sizes. For each simulated scenario, we generated 1,000 samples. In each sample, we recorded the false-negative rate of the five distribution-based methods with the best performance from the point of view of the false positives. Our results have revealed unacceptable rates of false negatives even with effects of very large size, starting from 31.8% in an optimistic scenario (effect size of 2.0 and a normal distribution) to 99.9% in the worst scenario (effect size of 0.2 and a highly skewed distribution). Therefore, our results suggest that the widely used distribution-based methods must be applied with caution in a clinical context, because they need huge effect sizes to detect a true change. However, we made some considerations regarding the effect size and the cut-off points commonly used which allow us to be more precise in our estimates.

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Viral Dynamics and Real-Time RT-PCR Ct Values Correlation with Disease Severity in COVID-19

Diagnostics ◽

10.3390/diagnostics11061091 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1091

Author(s):

Ali A. Rabaan ◽

Raghavendra Tirupathi ◽

Anupam A Sule ◽

Jehad Aldali ◽

Abbas Al Mutair ◽

...

Keyword(s):

Viral Load ◽

Real Time ◽

Disease Severity ◽

False Negative ◽

Influential Factors ◽

Confirmatory Test ◽

Acid Extraction ◽

Rt Pcr ◽

Viral Kinetics ◽

Ct Value

Real-time RT-PCR is considered the gold standard confirmatory test for coronavirus disease 2019 (COVID-19). However, many scientists disagree, and it is essential to understand that several factors and variables can cause a false-negative test. In this context, cycle threshold (Ct) values are being utilized to diagnose or predict SARS-CoV-2 infection. This practice has a significant clinical utility as Ct values can be correlated with the viral load. In addition, Ct values have a strong correlation with multiple haematological and biochemical markers. However, it is essential to consider that Ct values might be affected by pre-analytic, analytic, and post-analytical variables such as collection technique, specimen type, sampling time, viral kinetics, transport and storage conditions, nucleic acid extraction, viral RNA load, primer designing, real-time PCR efficiency, and Ct value determination method. Therefore, understanding the interpretation of Ct values and other influential factors could play a crucial role in interpreting viral load and disease severity. In several clinical studies consisting of small or large sample sizes, several discrepancies exist regarding a significant positive correlation between the Ct value and disease severity in COVID-19. In this context, a revised review of the literature has been conducted to fill the knowledge gaps regarding the correlations between Ct values and severity/fatality rates of patients with COVID-19. Various databases such as PubMed, Science Direct, Medline, Scopus, and Google Scholar were searched up to April 2021 by using keywords including “RT-PCR or viral load”, “SARS-CoV-2 and RT-PCR”, “Ct value and viral load”, “Ct value or COVID-19”. Research articles were extracted and selected independently by the authors and included in the present review based on their relevance to the study. The current narrative review explores the correlation of Ct values with mortality, disease progression, severity, and infectivity. We also discuss the factors that can affect these values, such as collection technique, type of swab, sampling method, etc.

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No evidence of false-negative Plasmodium falciparum rapid diagnostic results in Monrovia, Liberia

Malaria Journal ◽

10.1186/s12936-021-03774-3 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Mandella King ◽

Alexander E. George ◽

Pau Cisteró ◽

Christine K. Tarr-Attia ◽

Beatriz Arregui ◽

...

Keyword(s):

Plasmodium Falciparum ◽

False Negative ◽

Malaria Diagnosis ◽

Rapid Diagnostic Tests ◽

Molecular Testing ◽

False Negatives ◽

Gene Deletions ◽

History Of ◽

Catholic Hospital ◽

Or History

Abstract Background Malaria diagnosis in many malaria-endemic countries relies mainly on the use of rapid diagnostic tests (RDTs). The majority of commercial RDTs used in Africa detect the Plasmodium falciparum histidine-rich protein 2 (PfHRP2). pfhrp2/3 gene deletions can therefore lead to false-negative RDT results. This study aimed to evaluate the frequency of PCR-confirmed, false-negative P. falciparum RDT results in Monrovia, Liberia. Methods PfHRP2-based RDT (Paracheck Pf®) and microscopy results from 1038 individuals with fever or history of fever (n = 951) and pregnant women at first antenatal care (ANC) visit (n = 87) enrolled in the Saint Joseph’s Catholic Hospital (Monrovia) from March to July 2019 were used to assess the frequency of false-negative RDT results. True–false negatives were confirmed by detecting the presence of P. falciparum DNA by quantitative PCR in samples from individuals with discrepant RDT and microscopy results. Samples that were positive by 18S rRNA qPCR but negative by PfHRP2-RDT were subjected to multiplex qPCR assay for detection of pfhrp2 and pfhrp3. Results One-hundred and eighty-six (19.6%) and 200 (21.0%) of the 951 febrile participants had a P. falciparum-positive result by RDT and microscopy, respectively. Positivity rate increased with age and the reporting of joint pain, chills and shivers, vomiting and weakness, and decreased with the presence of coughs and nausea. The positivity rate at first ANC visit was 5.7% (n = 5) and 8% (n = 7) by RDT and microscopy, respectively. Out of 207 Plasmodium infections detected by microscopy, 22 (11%) were negative by RDT. qPCR confirmed absence of P. falciparum DNA in the 16 RDT-negative but microscopy-positive samples which were available for molecular testing. Among the 14 samples that were positive by qPCR but negative by RDT and microscopy, 3 only amplified pfldh, and among these 3 all were positive for pfhrp2 and pfhrp3. Conclusion There is no qPCR-confirmed evidence of false-negative RDT results due to pfhrp2/pfhrp3 deletions in this study conducted in Monrovia (Liberia). This indicates that these deletions are not expected to affect the performance of PfHRP2-based RDTs for the diagnosis of malaria in Liberia. Nevertheless, active surveillance for the emergence of PfHRP2 deletions is required.

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Need for Confirmatory Neutralization Tests for Hepatitis B Surface Antigen Tests in Populations with Intermediate Prevalence

Laboratory Medicine ◽

10.1093/labmed/lmab006 ◽

2021 ◽

Author(s):

Min Young Lee ◽

So Young Kang ◽

Woo In Lee ◽

Myeong Hee Kim

Keyword(s):

Hepatitis B ◽

Surface Antigen ◽

Neutralization Test ◽

Hepatitis B Surface Antigen ◽

False Negative ◽

Confirmatory Test ◽

Negative Group ◽

Positive Group ◽

Test Kit ◽

Antigen Tests

Abstract Objective Hepatitis B surface antigen (HBsAg) is known as the hallmark of hepatitis B virus (HBV) infection. This study aimed to determine whether an HBsAg neutralization test is necessary to accurately interpret HBsAg test results. Methods Initially reactive HBsAg specimens from a 5-year period, with cutoff index values between 1.0 and 2.0, were subjected to neutralization confirmatory testing using an Elecsys HBsAg Confirmatory test kit (Roche Diagnostics GmbH. Mannheim, Germany). Results The neutralization test showed 46.1% positive (confirmed positive group) and 53.9% negative (confirmed negative group) results from the total specimens. Among the confirmed negative group, 79.5% of patients were confirmed to be negative for the current infection, whereas 4 patients in the chronic hepatitis B subgroup showed a neutralization percentage close to 40%. More than half of patients in the confirmed positive group were considered to be in the hepatitis B e antigen-negative inactive HBsAg carrier phase. Conclusion In populations with intermediate HBV prevalence, a neutralization test is necessary to confirm an HBsAg result and reduce the false positive and false negative rates of initial HBsAg tests.

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Testing Segmentation Popular Loss and Variations in Three Multiclass Medical Imaging Problems

Journal of Imaging ◽

10.3390/jimaging7020016 ◽

2021 ◽

Vol 7 (2) ◽

pp. 16

Author(s):

Pedro Furtado

Keyword(s):

False Negative ◽

Magnetic Resonance Images ◽

Medical Image Segmentation ◽

Cross Entropy ◽

Loss Functions ◽

Optic Disk ◽

False Negatives ◽

Convolutional Network ◽

Percentage Points ◽

Class Background

Image structures are segmented automatically using deep learning (DL) for analysis and processing. The three most popular base loss functions are cross entropy (crossE), intersect-over-the-union (IoU), and dice. Which should be used, is it useful to consider simple variations, such as modifying formula coefficients? How do characteristics of different image structures influence scores? Taking three different medical image segmentation problems (segmentation of organs in magnetic resonance images (MRI), liver in computer tomography images (CT) and diabetic retinopathy lesions in eye fundus images (EFI)), we quantify loss functions and variations, as well as segmentation scores of different targets. We first describe the limitations of metrics, since loss is a metric, then we describe and test alternatives. Experimentally, we observed that DeeplabV3 outperforms UNet and fully convolutional network (FCN) in all datasets. Dice scored 1 to 6 percentage points (pp) higher than cross entropy over all datasets, IoU improved 0 to 3 pp. Varying formula coefficients improved scores, but the best choices depend on the dataset: compared to crossE, different false positive vs. false negative weights improved MRI by 12 pp, and assigning zero weight to background improved EFI by 6 pp. Multiclass segmentation scored higher than n-uniclass segmentation in MRI by 8 pp. EFI lesions score low compared to more constant structures (e.g., optic disk or even organs), but loss modifications improve those scores significantly 6 to 9 pp. Our conclusions are that dice is best, it is worth assigning 0 weight to class background and to test different weights on false positives and false negatives.

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Characteristics of patients who had a stroke not initially identified during emergency prehospital assessment: a systematic review

Emergency Medicine Journal ◽

10.1136/emermed-2020-209607 ◽

2021 ◽

pp. emermed-2020-209607

Author(s):

Stephanie P Jones ◽

Janet E Bray ◽

Josephine ME Gibson ◽

Graham McClelland ◽

Colette Miller ◽

...

Keyword(s):

Systematic Review ◽

Diagnostic Accuracy ◽

False Negative ◽

Visual Disturbance ◽

Screening Tools ◽

False Negatives ◽

Presenting Symptoms ◽

Ischaemic Attack ◽

Stroke Type ◽

Key Terms

BackgroundAround 25% of patients who had a stroke do not present with typical ‘face, arm, speech’ symptoms at onset, and are challenging for emergency medical services (EMS) to identify. The aim of this systematic review was to identify the characteristics of acute stroke presentations associated with inaccurate EMS identification (false negatives).MethodWe performed a systematic search of MEDLINE, EMBASE, CINAHL and PubMed from 1995 to August 2020 using key terms: stroke, EMS, paramedics, identification and assessment. Studies included: patients who had a stroke or patient records; ≥18 years; any stroke type; prehospital assessment undertaken by health professionals including paramedics or technicians; data reported on prehospital diagnostic accuracy and/or presenting symptoms. Data were extracted and study quality assessed by two researchers using the Quality Assessment of Diagnostic Accuracy Studies V.2 tool.ResultsOf 845 studies initially identified, 21 observational studies met the inclusion criteria. Of the 6934 stroke and Transient Ischaemic Attack patients included, there were 1774 (26%) false negative patients (range from 4 (2%) to 247 (52%)). Commonly documented symptoms in false negative cases were speech problems (n=107; 13%–28%), nausea/vomiting (n=94; 8%–38%), dizziness (n=86; 23%–27%), changes in mental status (n=51; 8%–25%) and visual disturbance/impairment (n=43; 13%–28%).ConclusionSpeech problems and posterior circulation symptoms were the most commonly documented symptoms among stroke presentations that were not correctly identified by EMS (false negatives). However, the addition of further symptoms to stroke screening tools requires valuation of subsequent sensitivity and specificity, training needs and possible overuse of high priority resources.

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