Quantification of Melanoma Micrometastases in Sentinel Lymph Nodes Using Real-Time RT-PCR

628 Background: Analysis using real time RT-PCR for the detection of metastases in lymph nodes (LN) increases sampling but is associated with a risk of too much sensitivity as compared to histological analysis. Determining the appropriate cutoff value is important in introducing this system to routine clinical practice. Our study confirms the reliability of the cutoff values of a real-time RT-PCR assay (GeneSearch, Veridex LLC) to detect metastases larger than 0.2 mm by detailed 0.2 mm frozen section histological diagnosis. Methods: 129 sentinel and non-sentinel lymph nodes were obtained by sentinel LN biopsy from 79 breast cancer patients. All LNs were cut in half. One half of each LN was used for routine intra-operative diagnosis. The other LN half had sections cut every 0.2 mm for histological analysis. All frozen sections were H&E stained and examined by on-site pathologists. These sections were then re-stained immunohistochemically using a pancytokeratin antibody (AE1/AE3) for detecting submicrometastses. All remaining tissue between the histology sections was assayed by the real-time RT-PCR assay using the genetic markers mammaglobin (MG) and cytokeratin-19 (CK-19) (note: tissue processing method was investigational and off-label). Cutoff values were pre-set in a large US study (n = 304). Results: Compared to the histological diagnosis using 0.2 mm interval frozen sections, the real-time RT-PCR results were as follows; sensitivity of 100.0% (34/34), specificity of 93.7% (89/95), and overall accuracy of 95.3% (123/129). Submicrometastases were recognized by IHC in 3 of 95 samples (2 solitary and 1 multiple sites) diagnosed as negative by H&E-stain sections, and 3 samples were negative for RT-PCR. Conclusions: Results of 0.2 mm interval histological analysis suggest a near perfect sensitivity of the real time RT-PCR assay, allowing reliable detection of LN metastases larger than 0.2 mm. In addition, at least three samples with submicrometastases detected by immunohistochemistry for pancytokeratin were above the preset cutoff values of this real time RT-PCR assay. [Table: see text]

Download Full-text

Assessing the status of axillary sentinel lymph nodes of breast carcinoma patients by a real-time quantitative RT-PCR assay for mammaglobin 1 mRNA

Breast Cancer Research and Treatment ◽

10.1007/s10549-005-9148-x ◽

2006 ◽

Vol 98 (2) ◽

pp. 185-190 ◽

Cited By ~ 14

Author(s):

Patrizia Dell’Orto ◽

Maria Olivia Biasi ◽

Barbara Del Curto ◽

Stefano Zurrida ◽

Viviana Galimberti ◽

...

Keyword(s):

Breast Carcinoma ◽

Lymph Nodes ◽

Real Time ◽

Sentinel Lymph Nodes ◽

Rt Pcr ◽

Pcr Assay ◽

The Status

Download Full-text

Detection of Micrometastases in Sentinel Lymph Nodes of Melanoma Patients by RT-PCR.

Nishi Nihon Hifuka ◽

10.2336/nishinihonhifu.64.211 ◽

2002 ◽

Vol 64 (2) ◽

pp. 211-217 ◽

Cited By ~ 1

Author(s):

Yoichi MOROI ◽

Chikage OBATA ◽

Shohei FUJITA ◽

Satoko SHIBATA ◽

Sawako KONISHI ◽

...

Keyword(s):

Lymph Nodes ◽

Sentinel Lymph Nodes ◽

Rt Pcr ◽

Melanoma Patients

Download Full-text

Reactive Lymphoid Hyperplasia or Tubercular Lymphadenitis: Can Real-Time PCR on Fine-Needle Aspirates Help Physicians in Concluding the Diagnosis?

Acta Cytologica ◽

10.1159/000488871 ◽

2018 ◽

Vol 62 (3) ◽

pp. 204-208 ◽

Cited By ~ 1

Author(s):

Vivek Gupta ◽

Arvind Bhake

Keyword(s):

Lymph Node ◽

Lymph Nodes ◽

Real Time ◽

Predictive Value ◽

Lymphoid Hyperplasia ◽

Reactive Lymphoid Hyperplasia ◽

Rt Pcr ◽

Likelihood Ratios ◽

Diagnostic Challenge ◽

Cross Sectional

Background: Enlarged lymph nodes in adult patients often present a diagnostic challenge. In the absence of granuloma or necrosis, the cytology/tissue findings are misleading and relate the enlarged lymph nodes to reactive lymphoid hyperplasia (RLH), because granuloma formation is an immunological response that usually takes 14–100 days to develop. This study assesses the role of real-time (RT)-PCR in the diagnosis of the Mycobacterium complex (MTBC) in lymph node aspirates compared with culture in cases of RLH. Methods: A cross-sectional study was conducted on 112 patients, aged 15–74 years, with a diagnosis of RLH on cytology. RT-PCR for MTBC detection and culture on Löwenstein-Jensen medium for tubercular bacilli was done on lymph node aspirates. Comparative values with reference to culture were calculated. The χ2 value, positive predictive value (PPV), negative predictive value (NPV), and likelihood ratios (LR) were calculated. Results: Out of 112 RLH cases, 35 (31%) were positive on both RT-PCR and culture. RT-PCR was positive in 43 cases and culture was positive in 44 cases. The χ2 test was found to be highly significant. PPV, NPV, positive LR, and negative LR were 81.4%, 87%, 6.76, and 0.23, respectively. Conclusion: RT-PCR for MTBC proves to be useful in arriving at a conclusive diagnosis in patients with a cytological diagnosis of RLH.

Download Full-text