Detection of Banana Mild Mosaic Virus in Musa In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species

: The banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects Musa spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four plants cultivated in a greenhouse for at least 6 months and causes a significant delay for the distribution of the germplasm. We evaluated the sensitivity of different protocols for BanMMV detection from in vitro plants to accelerate the testing process. We first used corm tissues from 137 in vitro plants and obtained a diagnostic sensitivity (DSE) of only 61% when testing four plants per accession. After thermotherapy was carried out to eliminate BanMMV infection, the meristem was recovered and further grown in vitro. The same protocol was evaluated in parallel on the corm tissue surrounding the meristem, as a rapid screening to evaluate virus therapy success, and was compared to the results obtained following the standard protocol. The obtained results showed 28% false negatives when conducting testing from corm tissues, making this protocol unsuitable in routine processes. Furthermore, RT-PCR and high-throughput sequencing (HTS) tests were applied on tissues from the base (n = 39) and the leaves (n = 36). For RT-PCR, the average DSE per sample reached 65% from either the base or leaves. HTS was applied on 36 samples and yielded 100% diagnostic specificity (DSP) and 100% DSE, whatever the sampled tissue, allowing the identification of a new Betaflexiviridae species infecting Musa. These results suggest that a reliable diagnostic of BanMMV from in vitro plants using RT-PCR or HTS technologies might represent an efficient alternative for testing after greenhouse cultivation.

Comparison of RNAscope and immunohistochemistry for evaluation of the UPK2 status in urothelial carcinoma tissues

Background UPK2 exhibits excellent specificity for urothelial carcinoma (UC). UPK2 evaluation can be useful in making the correct diagnosis of UC. However, UPK2 detection by immunohistochemistry (IHC) has relatively low sensitivity. This paper aimed to compare the diagnostic sensitivity of RNAscope and IHC for evaluation of the UPK2 status in UC. Methods Tissue blocks from 127 conventional bladder UCs, 45 variant bladder UCs, 24 upper tract UCs and 23 metastatic UCs were selected for this study. IHC and RNAscope were used to detect the UPK2 status in UCs. Then, comparisons of the two methods were undertaken. Results There was no significant difference between RNAscope and IHC for the evaluation of the UPK2 positivity rate in UC (68.0% vs. 62.6%, P = 0.141). Correlation analysis revealed a moderate positive correlation for detection of UPK2: RNAscope vs. IHC (P < 0.001, R = 0.441). Our results showed a trend toward a higher positive UPK2 rate detected by RNAscope (53.3%) than by IHC (35.6%) in variant bladder UCs. Disappointingly, the P value did not indicate a significant difference (P = 0.057). Conclusions RNAscope for UPK2 appeared to perform similarly to IHC, with a marginally higher positive rate, suggesting it could be used as an alternative or adjunct to UPK2 IHC.

Peroral Cholangioscopy-Guided Targeted Biopsy versus Conventional Endoscopic Transpapillary Forceps Biopsy for Biliary Stricture with Suspected Bile Duct Cancer

Background: The recent improvement of peroral cholangioscopy (POCS) maneuverability has enabled the precise, targeted biopsy of bile duct lesions under direct cholangioscopic vision. However, as only small-cup biopsy forceps can pass through the scope channel, the resulting small sample size may limit the pathological diagnosis of biopsy specimens. This study compared the diagnostic abilities of POCS-guided biopsy and conventional fluoroscopy-guided biopsy for bile duct cancer. Method: This multicenter, retrospective cohort study included patients exhibiting bile duct stricture with suspected cholangiocarcinoma in whom POCS-guided and fluoroscopy-guided biopsies were performed in the same session. The primary endpoint was the diagnostic sensitivity for malignancy. The size and quality of the biopsy specimens were also compared. Result: A total of 59 patients were enrolled. The sensitivity of POCS-guided biopsy was similar to that of fluoroscopy-guided biopsy (54.0% and 64.0%, respectively). However, when the modalities were combined, the sensitivity increased to 80.0%. The mean specimen size from POCS-guided biopsy was significantly smaller than that from fluoroscopy-guided biopsy. The specimen quality using fluoroscopy-guided biopsy was also better than that using POCS-guided biopsy. Conclusions: The diagnostic sensitivity of POCS-guided biopsy is still insufficient, mainly because of the limited specimen quantity and quality. Therefore, conventional fluoroscopy-guided biopsy would be helpful to improve diagnostic sensitivity.

The diagnostic value of contrast-enhanced ultrasound for cervical tuberculous lymphadenitis

OBJECTIVE: To investigate the value of contrast-enhanced ultrasound (CEUS) for the diagnosis of cervical tuberculous lymphadenitis (CTL). METHODS: The cohort study included 203 consecutive patients diagnosed with cervical lymph node. Before pathological or laboratory confirmation, all patients underwent CEUS examination, and the imaging findings were analyzed afterward. The diagnostic efficiency of the CEUS imaging findings of CTL was evaluated. RESULTS: Nighty-seven patients of the 203 (47.8%) were pathologically or laboratory confirmed with a CTL diagnosis while the remainder (52.2%) were diagnosed with non-tuberculous lymphadenitis. Regarding the imaging findings of CEUS, it was more common in CTL patients to find a pattern of heterogeneous enhancement inside the lymph nodes relative to non-tuberculous patients [81.44% (79/97) vs 15.09% (16/106), P < 0.01]. The sensitivity of the feature in diagnosis for CTL was 81.44% and the specificity was 84.91%, resepectively. Furthermore, a pattern of peripheral rim-like enhancement had been notable in CTL patients compared with non-tuberculous patients [86.60% (84/97) vs 12.26% (13/106), P < 0.01], associating with a diagnostic sensitivity of 86.60% and a specificity of 87.74% . When it came to the combination of both imaging findings mentioned above, the features were more prominent in CTL patients than compared with non-tuberculous patients [74.23% (72/97) vs 5.66% (6/106), P < 0.01], with a diagnostic sensitivity of 74.23% and a high specificity of 94.34% . Regarding area under curve (AUC) for the ROC analysis, the feature of internal heterogeneous enhancement, peripheral rim-like enhancement, and both features were 0.832, 0.872, and 0.843. CONCLUSIONS: CEUS patterns of heterogeneous enhancement and peripheral rim-like enhancement of lymph nodes are helpful characteristics for the diagnosis of CTL.

Diagnostic sensitivity of arginase, alpha-1 antitrypsin and alpha-fetoprotein in hepatitis patients

Chronic hepatitis C virus (HCV) and hepatocellular carcinoma (HCC) are common liver diseases that lead to death in Egypt, especially in men. The current study aimed to evaluate the diagnostic sensitivity of arginase (ARG) activity, alpha-1 antitrypsin (AAT), and alpha-fetoprotein (AFP) in the sera of patients with HCV (with & without viremia) and HCC. A total of 190 men classified as 40 healthy used as control (G1), 100 infected with HCV (subdivided into 50 with viremia (G2) and 50 without viremia (G3), and 50 with HCC (G4). The activity of ARG significantly decreased in HCV and HCC groups along with significant elevation in the level of AAT and AFP as compared with the control. Although a non-significant variation was scored in AST/ALT, significant differences were observed among AST/ARG and ARG/ALT in the pathogenic groups as compared with the healthy group. Moreover, significant variations in ARG, AAT, AFP, AST/ARG, and ARG/ALT were observed between viremia and non-viremia. Although AFP scored significant change among the viremia and HCC, the rest parameters scored non-significant changes between both groups. Furthermore, a receiver operating characteristic curve (ROC) showed the diagnostic ability for the selected parameters with high sensitivity and multiple linear regressions exhibited good associations between those parameters. These findings suggest the using possibility of ARG, AAT, and AFP in the diagnosis and/or follow-up of patients with HCV or HCC.