Lymph Node Aspirates Research Articles

Pitfalls in the Cytological Diagnosis of Nodal Hodgkin Lymphoma.

Hodgkin lymphoma (HL) is a hematopoietic neoplasm characterized by malignant Reed-Sternberg (RS) cells in an inflammatory background. Although the cytological features of HL are well elucidated in literature, yet many postulated factors cause its misdiagnosis. This study aims to assess the diagnostic reliability of fine needle aspiration cytology (FNAC) in HL and evaluate the factors contributing to a false-negative and false-positive diagnosis, taking histopathology as the gold standard. This was a retrospective study in which 47 cases of HL diagnosed on histopathology were compared with their prior cytological diagnosis. The patient's age ranged from 3 to 80 years (median: 36 years) with a M:F ratio of 2.9:1. Lymph node aspirations were performed from multiple anatomical sites, out of which the cervical was the most common (57.8%). FNAC was inconclusive in two cases due to unsatisfactory smears. The false-negative diagnosis of reactive lymphadenitis was given in four cases, and false-positive in four cases, which included three cases of non-HL, and one case of malignant small round blue cell tumor. The overall diagnostic accuracy of FNAC in the diagnosis of HL was 82.2%. The cytological diagnosis of HL can be challenging when classic RS cells are absent. Contributing factors for a false-negative diagnosis include obscuring reactive inflammatory cells, fibrosis of the involved lymph nodes, partial involvement of the lymph node, and misinterpretation. A thorough clinical examination with evaluation of FNAC smears from multiple areas, and ancillary tests help improve the diagnostic accuracy of cytological diagnosis.

Effects on lymph node size, staging and primary tumor histology on diagnostic accuracy of axillary lymph node aspirate of breast cancers.

Fine-needle aspiration cytology is preferred for axillary lymph node metastasis with low costs and minimal risks. To improve diagnostic performance by incorporating clinical-radiological-pathological parameters, a large cohort pre-operative aspirates in were reviewed for parameters affecting adequacy rate and accuracy. Axillary nodal aspirates from three institutions with histologic correlation were retrieved. Case notes were reviewed for parameters pertaining to the primary tumor, nodal status, histologic and cytologic diagnoses. Totally 1361 specimens were included. The risk of malignancy for C1-C5 categories were 53.39%, 27.45%, 70.97%, 83.33% and 88.00%, increasing to 75.86%, 94.59% and 99.28% for C3/C4/C5 categories excluding cases with neoadjuvant therapy. Node size (p < 0.001) and histologic grade (p = 0.003) of primary tumor positively correlated with specimen adequacy. Presence of in situ component trended towards inadequacy (p = 0.069). Lymph node size remained a strong predictor of concordant cytologic diagnosis (p < 0.001). A higher percentage of involved node (p = 0.006) and HER2 overexpressed breast cancers (p = 0.027) increased concordance. Cases with ≥ 4 (up to ≥ 10) positive nodes were more likely to be concordant (p = 0.009- < 0.001), with improvements of 8.27%-12.37%. For size, cut-offs of ≥ 5 and ≥ 10mm were significant (p = 0.006- < 0.001). It is critical that clinical-radiological-pathological findings be interpreted together with cytology. Aspirates from smaller nodes are more likely to be non-informative, irrespective of the total number of suspicious nodes, or a high-grade primary. In axillae with less than 4 suspicious nodes and/or a target node of less than 5-10mm, the diagnostic accuracy of aspiration cytology decreases and should be interpreted cautiously.

Acute myeloid leukemia with peripheral lymph node involvement in dogs: A retrospective study of 23 cases.

Acute myeloid leukemia (AML) can infiltrate extramedullary tissues, such as the liver, spleen, and lymph nodes and can be difficult to differentiate from lymphoma in cytologic and histologic specimens. Our goal was to identify cytologic features that would support a diagnosis of AML in peripheral lymph node aspirates, for which we used the term extramedullary AML (eAML). Medical records of 23 dogs with a diagnosis of AML and archived lymph node aspirate smears from 2016 to 2024 were reviewed across 4 institutions. Inclusion criteria included ≥50% myeloid blasts plus differentiating myeloid cells in lymph node smears, confirmation of myeloid lineage by flow cytometric analysis, and complete medical records. Peripheral lymphadenopathy was the reason for presentation (9/23, 39%) or was found incidentally on physical examination (14/23, 61%). Most dogs were bi- or pancytopenic (18/23, 78%), with blasts identified in blood smears of 18 dogs (78%). Initial lymph node aspirate interpretations included hematopoietic neoplasia (8/21, 38%), AML (6/21, 29%), lymphoma (5/21, 24%), lymphoid hyperplasia (1/21, 5%), and granulocytic precursor infiltrates (1/21, 5%). On lymph node smear review, cytologic features supporting an eAML were differentiating granulocytes, blasts with myeloid features or promonocytes, dysplastic changes in myeloid cells, and retention of residual lymphocytes. The median survival was 22 days (range = 1-360 days), and 69% of 16 dogs given chemotherapy or glucocorticoids lived for 30 days or more. Our study highlights the importance of hemogram results and lymph node aspirate smear examination for morphologic features of myeloid differentiation to help diagnose eAML in lymph node smears.

Detection and isolation of Leishmania infantum from natural infected dog in Türkiye

Some clinical signs such as cachexia, alopecia, exfoliative dermatitis, hair loss, and swollen lymph nodes were observed in a one-year-old crossbred male dog living in a rural area of Kirikkale province, Türkiye, was presented to the veterinary clinic by its owner. Anaemia, leucopenia, hyperglobulinemia, and bilirubinemia were detected. Seropositivity was detected using Leishmania IgG/IgM Rapid Test. The amastigote forms of the parasite were observed in the lymph node aspirates. Viable promastigotes were observed in the samples obtained from the Novy-MacNeal-Nicolle medium (NNN) tubes. The parasite DNA was extracted from the promastigotes produced in the NNN medium using a DNA extraction kit according to the manufacturer's instructions. The specific bands indicating the gene regions of L. infantum (350 bp and 730 bp for HSP20 and HSP70, respectively) were observed. Additionally, BLASTn analysis revealed 100 % similarity with GenBank-deposited L. infantum sequences. The sequences were submitted to GenBank (Accession numbers: OR806945 and OR806946). According to the author's knowledge, it is the first dog isolate deposited in the collection of the Microbiology Reference Laboratories and Biological Products Department, General Directorate of Public Health, Ministry of Health, Ankara, Türkiye. The isolate (TR_L. infantum CanL-1_damla) has been preserved by cryopreservation at −150 °C for further studies.

This is another ultrasound cine clip demonstrating a fine-needle aspiration of an axillary lymph node without (left) and with (right) steering.

This is another ultrasound cine clip demonstrating a fine-needle aspiration of an axillary lymph node without (left) and with (right) steering.

This ultrasound cine clip demonstrates a fine-needle aspiration of an axillary lymph node without (left) and with (right) steering.

This ultrasound cine clip demonstrates a fine-needle aspiration of an axillary lymph node without (left) and with (right) steering.

Tubercular dactylitis/Spina ventosa-A rare skeletal form of extrapulmonary tuberculosis.

A 9-year-old girl presented with progressive, painful swelling over the left index finger, associated with local signs of inflammation and restriction of finger movements. After the swelling was punctured using a needle, chronic discharging sinus formed. Examination revealed firm swelling over the right parotid area, bilateral cervical and left axillary lymphadenopathy, and firm swelling over the left index finger with a chronic discharging sinus over the lateral aspect of proximal phalanx. Mantoux intradermal injection was reactive (22x24 mm induration). Blood and pus cultures were sterile. Hand radiograph revealed dactylitis (Figure 1A-C). Fine aspiration and cytology of the cervical lymph node showed degenerated inflammatory cells, epithelioid granuloma and acid-fast bacilli. She was initiated on 4-drug antitubercular therapy (ATT). At 2-months follow-up, she was asymptomatic and had the pain and swelling over the left finger have reduced with significant radiological improvement. Tubercular dactylitis or 'spina ventosa' (wind-filled sail) refers to cystic expansion of short, tubular bones. It is commonly seen in younger children (between 1 and 6 years of age) and adults (between 20 and 50 years of age). A common presentation in endemic areas is presence of a painless, fusiform swelling over a single digit, most commonly the proximal phalanx of index or middle finger as in the index case, or over the metacarpals of middle and ring fingers. Earliest radiological clues include periosteitis followed by gradual destruction of bone and cyst formation in the cavity giving a ballooned-out appearance.

Mycobacterium terrae Complex Associated Lymphadenitis: Negative AFB PCR

Abstract Introduction/Objective Nontuberculous mycobacteria included with the Mycobacterium terrae complex (MTC) include a ubiquitous, slow-growing collection of mycobacteria known for causing severe resistant skin, joint, and bone infections. There are limited reports of M. Arupensis, included under MTC, causing human infections despite environmental ubiquity. Low sequencing sensitivity for species such as M. Arupensis may result in negative preliminary PCR speciation thus influencing the negative predictive value of this method of organism detection. No prior case reports have described MTC infection presenting as subacute lymphadenitis. Methods/Case Report A 48 year old El Salvadorian male with known diagnosis of HIV presented with oral thrush and significant left-sided lymphadenopathy. He was initially diagnosed with HIV in El Salvador and on ART until immigrating to the US six months prior. HIV antigen and antibody was positive with positive reflex viral PCR. QuantiFERON-TB Gold resulted positive, but CXR was without evidence of pulmonary TB. TB1, TB2, and mitogen minus NIL were negative. He was referred for fine needle aspiration and core needle biopsy of the left cervical lymph node. AFB and AFB FITE stains were performed, but no definitive mycobacteria were identified. GMS was performed and negative for fungal elements. A final histological diagnosis of necrotizing granuloma was given and the tissue was sent for bacterial, AFB, and fungal DNA PCR; no organisms were detected. A sputum sample was obtained and cultured four months after initial presentation and was positive for acid fast bacilli: Mycobacterium terrae complex, most closely resembling Mycobacterium arupensis. The Fite and AFB stains were again reviewed, revealing one single acid fast bacilli organism identified on Fite stain in the necrotizing granuloma. Results (if a Case Study enter NA) N/A Conclusion The negative AFB DNA PCR result may have been a false negative result due to poor sensitivity or selectivity for M. arupensis. There are no former reported cases of M. Arupensis associated necrotizing granulomas in lymph node. This unusual MTC infection presenting as chronic unilateral lymphadenitis underscores the importance of broadening clinical laboratory AFB DNA PCR sensitivity especially in cases of suspected AFB or modified AFB associated necrotizing granulomas.

Saurian-associated Leishmania tarentolae in dogs: Infectivity and immunogenicity evaluation in the canine model.

In canine leishmaniosis endemic areas, Leishmania infantum may occur in sympatry with the non-pathogenic Leishmania tarentolae, which is associated to reptiles. The potential infectivity of L. tarentolae for mammals raises questions about the interactions between the two Leishmania species, and the potential cross-immune protection in dogs. This study aimed to assess the outcome of experimental L. tarentolae infection in dogs, determining: i) the anti-L. tarentolae antibody production, ii) the duration of the immunity and cytokine expression, and iii) the possible pathogenic effect in the canine host. Twelve purpose-bred beagle dogs were randomly allocated to three groups (intravenous inoculation, G1; intradermal inoculation, G2; negative control, G3). G1 and G2 dogs were inoculated twice (day 0, day 28) with 108 promastigotes of L. tarentolae strain (RTAR/IT/21/RI-325) isolated from a Tarentola mauritanica gecko. The animals were followed until day 206. Blood, serum, conjunctival swabs and lymph node aspirate samples were collected monthly and bone marrow, liver and spleen biopsies on day 91. Hematological and biochemical parameters were assessed monthly, as well as serology (IFAT and ELISA) and molecular identification of L. tarentolae. Mononuclear cells (PBMC) were obtained to assess the cytokine expression through in vitro stimulation or (re-) infection. Data from this study demonstrated that DNA from L. tarentolae is detectable up to 3 months post-infection, with seroconversion after day 28. Moreover, the non-pathogenic nature of L. tarentolae was confirmed, with a neutral Th1/Th2 polarization, and a possible shift to Th1 phenotype after derived macrophages (re-) infection, as demonstrated by the expression of IFN-gamma. Therefore, L. tarentolae demonstrated a great potential as a surrogate pathogen and/or immune-prophylaxis/immune-therapy against Leishmania infections in dogs and humans.

Cartridge-Based Nucleic Acid Amplification Test Compared to Fine Needle Aspiration Cytology in Suspected Cases of Tubercular Lymphadenitis (the Indian Experience)

INTRODUCTION: Diagnosis of tubercular lymphadenitis is daunting as there are varied clinical presentations and no single confirmatory gold standard test. Cartridge-based nucleic acid amplification test (CBNAAT) of the lymph node is a rapid molecular diagnostic test for simultaneously detecting tuberculosis (TB) and rifampicin resistance. AIM: To evaluate the performance of the CBNAAT test for detecting M. tuberculosis in lymph node specimens compared to fine needle aspiration cytology (FNAC). MATERIALS AND METHODS: The study was conducted in a rural tertiary care hospital in central India. A total of 180 patients clinically suspected of tubercular lymphadenitis were included. The male-to-female ratio was 1:1.3. The average age was 33.3 years. The age group 21–40 years had the highest number of cases. The most common complaints among the patients were fever (29.4%), followed by loss of appetite (9.5%), weight loss (9.5%), and cough (6.6%). However, most patients presented to the hospital with only lymphadenopathy (44.4%). The most common site involved was the anterior cervical lymph node (78.8%), followed by the axillary group (10.5%), submandibular (2.8%), inguinal (2.8%), supraclavicular (2.2%), submental (1.7%) and infraclavicular (1.1%) group of lymph nodes. The patients were subjected to both FNAC and CBNAAT testing. Results were reported as positive or negative for M. tuberculosis as CBNAAT gives a semiquantitative estimate of the concentration of bacilli. Rifampicin resistance results were reported as detected or not detected. RESULTS: Cytological examination of the lymph node aspirates revealed that most were tubercular lymphadenitis cases. Cytomorphological analysis of the cases of tubercular lymphadenitis revealed Type 6 (tubercular abscess) as the predominant pattern. CBNAAT testing detected 26 cases of M. tuberculosis and three cases of rifampicin resistance. The study reported a specificity of 92.92% and low sensitivity of 26.86% of combined FNAC and CBNAAT is much higher compared to only CBNAAT. CONCLUSION: CBNAAT, along with FNAC, is a valuable addition in first-line investigations of tubercular lymphadenitis to make a timely diagnosis.

A-104 Study of the matrix effect for thyroglobulin and calcitonin in fine-needle aspiration of cervical nodules

Abstract Background Analysis of material from fine needle aspiration of cervical thyroglobulin nodules with Beckman Coulter reagents on the ACCESS equipment and Calcitonin using the Siemens reagent on the Immulite equipment. The matrix effect, linearity and the limit of analytical sensitivity were analyzed. This study aims to analyze the correlation between the serum/plasma results described in the suppliers' package inserts with the results obtained with the puncture matrix. Methods The performance characteristics were tested according to the laboratory's operating procedures, based on the recommendations of ANVISA RDC 166/2017. Such parameters involved doping, which corresponds to the introduction of a calibrator with a known and traceable value into a pool of puncture material that is known to be negative, in order to analyze the behavior of the results obtained, paying attention to the deviations that may eventually occur due to the interference of the characteristics of the puncture material. puncture. After doping the material, dilutions were performed up to the sensitivity limit of the test, also analyzing the functional sensitivity, where ten replicates of the material were dosed, thus obtaining CV%, mean and Standard Deviation values. The same process was carried out and evaluated using the serum material. During the entire process, the materials received and analyzed were centrifuged and always kept in a refrigerated environment. Results The recovery% of the matrix effect study for cervical aspirate puncture showed results within the expected range, ranging from 80% to 120%, as well as the results compared with serum/plasma referenced in the supplier's package insert. The functional sensitivity study for Thyroglobulin showed a CV% of 1.77%, being lower than that recommended by the package insert (2.2%) and for the Calcitonin analyte, the functional sensitivity study showed a CV% of 8.49%, being lower than recommended by the package leaflet (15.7%), in addition to having CV's lower than the 20% recommended by ANVISA. The results obtained with the matrix effect study for serum also showed values between 80% and 120%, both of which agreed. Conclusions The results showed excellent performance for material from fine needle aspiration for both Calcitonin and Thyroglobulin, being consistent with the serum data described by the suppliers' package inserts, thus making the reagent usable for the cervical aspirate puncture matrix.

Primary mast cell tumour of the soft palate in a dog

AbstractA 12‐year‐old, male, entire Labrador retriever was presented to its primary care practice with a history of vomiting. A soft tissue mass was identified within the soft palate. Cytology from a fine‐needle aspirate revealed atypical mast cells, consistent with a mast cell tumour. Initial staging included complete blood count, serum biochemistry, serum electrolyte assay, thoracic, abdominal and cervical radiographs, computed tomography of the head and neck, and ultrasound‐guided aspirates of the retropharyngeal lymph nodes, liver and spleen. No evidence of nodal or distant metastasis was identified, and a diagnosis of primary soft palatal mast cell tumour was made. The dog underwent staphylectomy followed by adjunctive chemotherapy. Repeat staging was performed, which did not identify any evidence of metastasis. The dog died 22 months after initial presentation from an unknown cause.

Cytological spectrum of malignant lesions of lymph node

Background: Fine needle aspiration cytology (FNAC) is a safe, simple, non-invasive and inexpensive procedure for diagnosing the lesions of lymph nodes. It helps in the diagnosis of benign, infectious lesions as well as malignant lesions. It not only confirms or excludes metastasis in a case of a known primary malignancy but, also, helps to detect occult primary malignancy. Aim of the present study was to determine the frequency of malignant lesions of lymph nodes in FNAC. Methods: Retrospective study included all cervical lymph node lesions reported on FNAC, in the department of pathology, NSCB Medical College, Jabalpur from January 2023 to June 2024 (18 months). Results: Total cases of 414 lymph node aspiration were done, of which 78 cases (18.84%) were positive for malignancy. Among malignant lesion metastasis was the predominant lesion. Majority of cases were seen in 51-60 years age group with male predominance and male: female ratio was 6.1:1. The most common metastasis was from squamous cell carcinoma which was reported in 65 cases (84%), followed by 4 cases of adenocarcinoma and malignancy of epithelial origin each. One case each of metastasis from papillary carcinoma thyroid, anaplastic carcinoma thyroid and malignant melanoma was reported. In two cases cytological diagnosis of lymphoma (NHL) was made. Conclusions: FNAC gives early and accurate results with minimal invasion and reduces the need for surgical biopsies, thus saves cost and time to reach the final diagnosis. It is concluded that FNAC is a useful tool in diagnosing malignant lesions of lymph nodes with a good certainty.

Adequacy Assessment in Lymph Node Aspirates: An Exploratory Cytomorphologic Analysis of Negative Cervical Node Aspirates of Head and Neck Carcinomas

Introduction: Fine-needle aspiration cytology (FNAC) of lymph node is sensitive for detection of metastatic carcinoma but not without a significant false-negative rate. This study reviews clinicocytological features of negative node aspirates to identify predictive factors for establishing adequacy criteria. Methods: Negative FNAC specimens matched with neck dissection from a primary diagnosis of head and neck squamous cell, or undifferentiated (nasopharyngeal) carcinoma were reviewed for clinical and cytological parameters including lymphoid, inflammatory, and background components. Results: Slides from 86 lymph node aspirates including 50 positive for metastasis on follow-up were retrieved. Higher total lymphocyte count, lymphoid fragment count, germinal center fragment count, undifferentiated histology, presence of histiocytes and absence of blood were associated with a true negative cytologic diagnosis (p < 0.05), but not node size or location (p > 0.05). Undifferentiated histology, small lymphoid and germinal center fragments were independent factors indicative of a true negative diagnosis (p < 0.05). Large lymphoid fragments (p = 0.052) demonstrated a trend. Assessment of lymphoid components over five hotspots high-power fields (HPFs) was more robust in predictive value than only one hotspot. Receiver operating characteristic curve identified >10 small lymphoid, >20 large lymphoid and >2 germinal center fragment per five HPFs as optimal adequacy thresholds. Stricter total lymphocyte count cutoff accompanies increase of diagnostic accuracy, up to 0.67 for ≥5 HPFs with >500 lymphocytes. Conclusion: Total counts of lymphoid and germinal center fragments from multiple HPFs are useful in adequacy assessment of lymph node aspirates and improve diagnostic performance of FNAC in exclusion of metastatic carcinoma.

Cholangiocarcinoma: Consistent clinical, cytological, hematological, and biochemical findings and pathomorphology of the liver and kidney in five exotic dog breeds in Abeokuta, Nigeria.

Cholangiocarcinomas are malignant neoplasms that originate from any part of the bile duct epithelium. It is one of the most common liver tumors in dogs. This study described the clinical, cytological, hematological, biochemical, and pathomorphological findings of five cholangiocarcinoma cases in exotic breed dogs aged 2-5 years to aid in clinical diagnosis. This study used dogs presented at different times from 2012 to 2021 at the Veterinary Teaching Hospital, Federal University of Agriculture, Abeokuta. History, clinical signs, and vital parameters were recorded. Blood samples were collected for hematology and serum chemistry. Abdominocentesis was performed for cytological diagnosis. All dogs died during treatment, and postmortem examinations were performed. At postmortem, fine needle aspirates were collected from the liver and mesenteric lymph nodes and liver and kidney samples were fixed in 10% neutral-buffered formalin. The dogs showed signs of severe malnutrition, jaundice, and bloating. The hematological analysis indicated anemia, neutrophilia without band neutrophils, and lymphopenia, indicative of a stress hemogram. The serum biochemistry test revealed lower levels of total proteins, albumin, and globulin and higher levels of serum enzymes. Abdominal fluid and mesenteric lymph node cytology revealed clusters of epithelial neoplastic cells. A postmortem examination revealed the liver's nodular enlargement with the presence of button-like ulcers. Neoplastic epithelial cells are solid masses with hyperchromatic nuclei surrounded by fibrous connective tissues. Cholangiocarcinoma, diagnosed over a period of time in five exotic breeds of dog, consistently presents with the same clinical and postmortem findings, aiding in clinical diagnosis. However, the diagnosis of the disease is not possible in the early stage because of the absence of specific clinical signs. In dogs and possibly other animal species presenting with emaciation, lethargy, icterus, and distended abdomen, cholangiocarcinoma should be suspected, and cytological examination of the abdominal fluid and lymph node aspirates should be performed despite the absence of advanced equipment.

Sydney Reporting System for Lymph Node Fine-Needle Aspiration and Malignancy Risk Stratification: Is It of Clinical Value?

Lymphadenopathy is a common presentation of both reactive and malignant diseases, and lymph node fine-needle aspiration cytology (LN-FNAC) is an effective and inexpensive screening method. It can prevent unnecessary invasive surgery and excisional biopsy, especially in benign cases. Unfortunately, the lack of universally accepted terminology for reporting results has hindered its widespread support. The Sydney system proposal for lymph node cytopathology categorization and reporting introduced five diagnostic categories to address the lack of universally accepted terminology for reporting results in lymphadenopathy. Our study analyzed 188 lymph node fine-needle cytology (FNC) samples from King Abdulaziz University Hospital, Saudi Arabia, examining clinical follow-up data, pathology records, patient information, and final diagnosis from January 2019 to December 2022. Most specimens were from axillary lymph nodes, with 99.5% tissue correlation. The Sydney system category classification identified 56.9% of cases as malignant, while 26.1% were benign. The final surgical specimen diagnosis revealed a higher percentage of malignant diagnoses, with the highest risk of malignancy (ROM) in malignant/category V. In conclusion, our study demonstrates that LN-FNAC offers high diagnostic accuracy for lymph node (LN) aspirates, with the Sydney approach potentially aiding risk stratification and achieving consistency in cytologic diagnosis, but further multi-centric research is needed.

Feasibility of Using Needle Rinse Fluid for Cobas Human Papillomavirus (HPV) Assay in Diagnosing HPV+ Oropharyngeal Cancer with Neck Lymph Node Aspiration.

Human papillomavirus (HPV) is a crucial prognostic factor in oropharyngeal cancer (OPC). p16 is a surrogate marker for diagnosing HPV+ OPC, however it is not direct evidence of HPV existence. The purpose of our study was to evaluate an HPV DNA test-Cobas HPV assay-in diagnosing HPV+ OPC through neck lymph node aspiration. Patients with suspected neck mass who received fine needle aspiration (FNA) or core needle biopsy (CNB) at the National Taiwan University Hospital between January 2018 and December 2022 were reviewed. Besides routine cytology and pathology study, needle rinse fluid was collected for the Cobas HPV assay to detect high-risk HPV. We analyzed 137 patients with suspected lymph nodes, 32 (23.4%) of whom were HPV+ OPC patients and 105 (76.6%) of whom had non-HPV-related disease. FNA was performed in 31 patients and CNB was performed in 106 patients, according to the size and necrosis status of the lymph nodes. For diagnosing HPV+ OPC, CNB combined with p16 immunohistochemistry staining showed sensitivity of 93.3%, specificity of 97.8%, positive predictive value (PPV) of 87.5%, negative predictive value (NPV) of 98.9%, and accuracy of 97.2%. On the other hand, for the needle rinse Roche Cobas HPV assay, the test showed sensitivity of 96.9%, specificity of 100%, PPV of 100%, NPV of 99.1%, and accuracy of 99.3%. Compared with p16 IHC staining, the Cobas HPV test showed better PPV with statistical significance (p=0.04). The Cobas HPV assay is a US FDA-approved, highly automated, and readily used technique to directly detect the presence of high-risk HPV. We recommend utilizing the Cobas HPV assay in combination with routine cytology or histopathology examination in the work-up of neck lymphadenopathy.

Clinical manifestations and diagnostic approaches in cases of canine leishmaniasis in Bulgaria.

Leishmaniasis, a parasitic disease found in many parts of southern Europe, is transmitted in both humans and canines through the bite of phlebotomine sandflies, and can present in a variety of ways, such as cutaneous, mucocutaneous, diffuse, and visceral. In Bulgaria there are endemic areas of canine leishmaniasis, with sporadic cases in humans. However, no detailed studies of the animal population and vectors have been performed. Here we describe a few clinical cases of canine visceral leishmaniasis in two districts in western Bulgaria: one endemic and one without previously detected cases in humans or dogs. Diagnosis was confirmed serologically and molecularly using both real time and conventional PCR. Specific anti-leishmanial antibodies were confirmed in three of the cases via ELISA, with 50% of them returning extremely high values. In the majority of the cases DNA fragments were detected in the skin or lymph node aspirate but not in the blood. This paper highlights the need for further studies updating the current knowledge on the epidemiology, diagnosis, and control of visceral leishmaniasis in the reservoir host population.

Multi-centric evaluation of Truenat MTB and MTB-RIF Dx assays for diagnosis of extrapulmonary tuberculosis

Extra-pulmonary TB (EPTB) is difficult to diagnose due to paucibacillary nature of disease. Current study evaluated accuracy of Truenat MTB and MTB-Rif Dx (TN), for detection of Mycobacterium tuberculosis and resistance to rifampicin. Samples were collected from 2103 treatment naive adults with presumptive EPTB, and tested by smear microscopy, liquid culture (LC) (MGIT-960) and GeneXpert MTB/RIF (GX) (Microbiological Reference Standards, MRS). TN results were compared to MRS and Composite Reference Standards (CRS, Microbiology, histopathology, radiology, clinical features prompting decision to treat, response to treatment). CRS grouped patients into 551 confirmed, 1096 unconfirmed, and 409 as unlikely TB. TN sensitivity and specificity was 73.7% and 90.4% against GX. Against LC, Overall sensitivity of GX was 67.6%, while that of TN was 62.3%. Highest sensitivity by TN was observed in pus samples (89%) and highest specificity (92%) in CSF samples, similar to GX. TN sensitivity was better in fluid and biopsy samples and slightly inferior for lymph node aspirates compared to GX. TN sensitivity for RIF resistance detection was slightly superior to GX. TN and GX results were further compared to Clinical Reference Standards. TN detected 170 TB patients initiated on treatment missed by GX, while GX detected 113 such patients missed by TN. Of 124 samples with RIF resistance discordance between GX and TN, GX reported 103/124 as sensitive, 3/124 as indeterminate and 18 as resistant (13/18 samples had low/very low DNA load) while TN reported RIF resistance indeterminate in 103/111 low/very low DNA load samples. Due to paucibacillary nature of EPTB samples, culture yield was poor and phenotypic drug susceptibility testing failed to resolve the discordance. The study establishes TN at par with GX and can be utilized for quick and accurate diagnosis of EPTB.

Insights into changing patterns of extrapulmonary tuberculosis in North India

PurposeTuberculosis is one of the dreadful infections and India contributes to substantial burden of TB cases globally. Though majority of cases are pulmonary, extra-pulmonary tuberculosis (EPTB) share significant burden, more in HIV-positive persons. Despite the striking burden, very few studies have been conducted in India and present study was undertaken to determine trends of EPTB at our tertiary care centre. MethodsThis was a retrospective study conducted over a period of 4 years 3 months. Diagnosis of EPTB was based on suspected clinical features, with positive micobiological evidence with cartridge based nucleic acid amplification test (CBNAAT) with/without microscopy. ResultsA total of 10,560 samples (pulmonary and extra-pulmonary) were received during the study period, of which 3972 were extrapulmonary. Of these, a total of 18% were noted to be positive for EPTB. Trend of positivity revealed highest burden in 2018 and a decline was noted over the years, however, rise in cases was noted in 2022. Pleural, meningitis, musculoskeletal, peritoneal and pericardial TB was more common in males, while lymphadenitis was more common in females (p value: <0.0001). Pleural TB (31%) was the most common presentation, followed by lymphadenitis. A gradual decline in lymphadenitis was noted with significantly increasing trend only for musculoskeletal TB. Rifampicin resistance was detected in 7.45% of positive samples, of which the maximum rate of resistance was noted in lymph node aspirates (11.11%), followed by musculoskeletal and pleural samples. ConclusionThe present study showed a gradual decline in positivity of EPTB cases over the years. Younger productive age group with more propensity to transmit infection was the most commonly affected, with pleural TB as the most common presentation. Rare presentations of EPTB also contributed major share. Higher rates of resistance underline requisite to strengthen ongoing programs, to achieve the End TB strategy by 2025.