Rapid and Specific Diagnosis of Extrapulmonary Tuberculosis by Immunostaining of Tissues and Aspirates With Anti-MPT64

Purohit, Manju; Sviland, Lisbeth; Wiker, Harald G.; Mustafa, Tehmina

doi:10.1097/pai.0000000000000300

Cited by 30 publications

(26 citation statements)

References 17 publications

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“…When a diagnostic validation of immunostaining was performed using PCR as the gold standard, the overall sensitivity, specificity, and positive and negative predictive values for immunostaining with anti-MPT64 were 100%, 97%, 98%, and 100%, respectively. (21) IHC for tuberculosis has, however, been slow to catch on as a routine diagnostic method in histopathology laboratories probably due to the lack of a specific anti-mycobacterial antibody suitable for all types of tissues and hence the exact diagnostic role of IHC for M. tuberculosis has to be assessed. Present study shows that IHC with anti-MPT64 has better specificity, sensitivity, and predictive values.…”

Section: Discussionmentioning

confidence: 99%

Study of CBNAAT and Anti-MPT64 Detection in Cytological and Histopathological Material for Early Diagnosis of TB Lymphadenitis

Sumayya¹,

Kharidehal²,

S³

et al. 2019

jemds

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BACKGROUND Tuberculosis is primarily a pulmonary disease, but it can affect almost any organ system, with lymph node involvement being the most common form of extrapulmonary tuberculosis and tuberculous pleuritis being the 2 nd most frequent extrapulmonary manifestation. Immune-compromising diseases such as HIV, have resulted in increase in the incidence of tuberculous lymphadenitis. METHODS This study is a prospective study, which included 80 patients, in the age group of 15-60 years, done over a period of 18 months from November 2016 to April 2018 in Upgraded Department of Pathology, Osmania General Hospital, Hyderabad, a tertiary referral centre for the state of Telangana and various districts of neighbouring states. In the present study, patients who presented with cervical lymphadenopathy as the manifestation of extrapulmonary tuberculosis were included. Cervical lymphadenopathy is the most common presentation of EPTB. Pus from the suspected lymph nodes was aspirated for cytological examination, ZN staining and GeneXpert analysis. Biopsy from the suspected lymph node was obtained and the material processed for CBNAAT, histopathology, Ziehl Neelsen staining and IHC. RESULTS CBNAAT was positive in 70 (87.5%) cases on cytological aspirates with 24 (30%) cases showing rifampicin resistance and on biopsy, 64/80 (80%) cases were positive on CBNAAT with rifampicin resistance in 25 (31.25%) cases. IHC with anti-MPT64 antibody was positive in 72 (90%) cases with sensitivity, specificity, positive and negative predictive value of 100%, 97%, 98% and 100% respectively and p value of <0.0001. CONCLUSIONS Present study provides the insight for the feasibility of using CBNAAT and IHC as diagnostic techniques for better management of EPTB patients and be useful in settings where culture or higher molecular facilities are not available.

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Section: Discussionmentioning

confidence: 99%

Study of CBNAAT and Anti-MPT64 Detection in Cytological and Histopathological Material for Early Diagnosis of TB Lymphadenitis

Sumayya¹,

Kharidehal²,

S³

et al. 2019

jemds

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“…Rapid and accurate TB diagnosis using pathogen markers remains a difficult goal. Detection of MPT64 and antigen 85B via immunostaining of tissues and fine-needle aspirates can be rapid and sensitive methods for establishing an early and specific diagnosis of TB infection but are limited to pathology and microbiology laboratories ( 42 , 43 ). Detection of MPT64 in sputum cultures utilizing an immune-chromatographic assay, in conjunction with cording in a stained smear, has been used to make a preliminary yet high-confidence identification of M. tuberculosis complex in liquid culture ( 44 , 45 ).…”

Section: Discussionmentioning

confidence: 99%

Potential of High-Affinity, Slow Off-Rate Modified Aptamer Reagents for Mycobacterium tuberculosis Proteins as Tools for Infection Models and Diagnostic Applications

et al. 2017

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Direct pathogen detection in blood to diagnose active tuberculosis (TB) has been difficult due to low levels of circulating antigens or due to the lack of specific, high-affinity binding reagents and reliable assays with adequate sensitivity. We sought to determine whether slow off-rate modified aptamer (SOMAmer) reagents with subnanomolar affinity for Mycobacterium tuberculosis proteins (antigens 85A, 85B, 85C, GroES, GroEL2, DnaK, CFP10, KAD, CFP2, RplL, and Tpx) could be useful to diagnose tuberculosis. When incorporated into the multiplexed, array-based proteomic SOMAscan assay, limits of detection reached the subpicomolar range in 40% serum. Binding to native M. tuberculosis proteins was confirmed by using M. tuberculosis culture filtrate proteins and fractions from infected macrophages and via affinity capture assays and subsequent mass spectrometry. Comparison of serum from culture-positive pulmonary TB patients and TB suspects systematically ruled out for TB revealed small but statistically significant (P < 0.0001) differences in the median M. tuberculosis signals and in specific pathogen markers, such as antigen 85B. Samples where many M. tuberculosis aptamers produced high signals were rare exceptions. In concentrated, protein-normalized urine from TB patients and non-TB controls, the CFP10 (EsxB) SOMAmer yielded the most significant differential signals (P < 0.0276), particularly in TB patients with HIV coinfection. In conclusion, direct M. tuberculosis antigen detection proved difficult even with a sensitive method such as SOMAscan, likely due to their very low, subpicomolar abundance. The observed differences between cases and controls had limited diagnostic utility in serum and urine, but further evaluation of M. tuberculosis SOMAmers using other platforms and sample types is warranted.

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“…MPT64 detection had the best performance in TB lymphadenitis patients and children with TB, suggesting that MPT64 detection could be used for routine diagnosis under low resource allocation to improve the diagnosis of extrapulmonary tuberculosis, especially for tuberculous lymphadenitis and children with TB [39] . Purohit et al studied 51 cases of pulmonary tuberculosis and 38 control specimens of non-pulmonary tuberculosis.The results suggested that anti-MPT64 immunostaining detection was a rapid and sensitive method for early specific diagnosis of Mycobacterium tuberculosis infection [40] . This technique can easily be incorporated into routine pathological laboratories.…”

Section: Detection Of Mtbc By Intestinal Tuberculosis Protein Combinementioning

confidence: 91%

Application value of tissue tuberculosis antigen combined with Xpert MTB/RIF detection in differential diagnosis of intestinal tuberculosis and Crohn’s disease

Fei

Zhou

Luo

et al. 2020

Preprint

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Background: The purpose of this study was to evaluate the value of Xpert MTB/RIF detection and tuberculosis antigen detection of Mycobacterium tuberculosis cluster (MTBC) in intestinal tissues for differentiating intestinal tuberculosis (ITB) from Crohn’s disease (CD). Methods: A total of 110 patients who were clinically diagnosed with CD or ITB were monitored. Several specimens of intestinal tissue from endoscopic biopsy or surgical excision were used for culture and Xpert MTB/RIF for detection of MTBC, respectively. Four antigens (38KDa, ESAT-6, MPT64, Ag85 complex) of MTBC in intestinal tissue were detected by immunohistochemistry. Results: A total of 42 cases of intestinal tuberculosis and 46 cases of CD were included in the experimental analysis. Perianal lesions and longitudinal ulcers were more common in CD patients (p < 0.05), while caseous granuloma and annular ulcers were more common in ITB patients (P < 0.05). The positive rate of MTBC detected by Xpert MTB/RIF in intestinal tissue samples of ITB patients was 33.33%, which was significantly higher compared to CD patients (p < 0.05) and compared to acid-fast staining smears (9.52%) (p < 0.05). The positive MPT64 expression rate in patients with intestinal tuberculosis was 40.48%, which was significantly higher than that observed in CD patients, which was 19.56% (p<0.05). Conclusions: The detection of Xpert MTB/RIF in intestinal tissue is a rapid and useful method for establishing an early diagnosis of intestinal tuberculosis. The detection of Xpert MTB/RIF and MPT64 antigen in intestinal tissues have definitive value in the differential diagnosis of intestinal tuberculosis and Crohn’s disease. The combination of these two methods could improve detection sensitivity.

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Rapid and Specific Diagnosis of Extrapulmonary Tuberculosis by Immunostaining of Tissues and Aspirates With Anti-MPT64

Cited by 30 publications

References 17 publications

Study of CBNAAT and Anti-MPT64 Detection in Cytological and Histopathological Material for Early Diagnosis of TB Lymphadenitis

Study of CBNAAT and Anti-MPT64 Detection in Cytological and Histopathological Material for Early Diagnosis of TB Lymphadenitis

Potential of High-Affinity, Slow Off-Rate Modified Aptamer Reagents for Mycobacterium tuberculosis Proteins as Tools for Infection Models and Diagnostic Applications

Application value of tissue tuberculosis antigen combined with Xpert MTB/RIF detection in differential diagnosis of intestinal tuberculosis and Crohn’s disease

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