乳腺癌前哨淋巴结术中分子诊断的临床实用性初探

Objective To evaluate the application of GeneSearchTM breast lymph node assay in intraoperative detection of metastases in sentinel lymph node (SLN) from breast cancer patients. Methods A total of 225 SLN from 88 patients was prospectively studied. Each SLN was cut into 2 mm slabs which were examined by intraoperative imprint cytology (ⅡC) first, followed by GeneSearch assay and postoperative serial sectioning. GeneSearch used real-time fluorescence quantitative RT-PCR technology to detect the expression of CK19 and mammaglobin in SLN. The results of GeneSearch assay were correlated with those of ⅡC and post-operative serial sectioning. Results Amongst the 88 cases studied, 225 SLNs were found, and obvious metastatic carcinoma cells were identified in 27 SLNs and micrometastasis in 9 SLNs.One hundred and eight-nine SLNs were considered as "negative" (with "isolated tumor cells" present in 5 SLNs). The turn-around time of intraoperative GeneSearch assay ranged from 35 to 45 minutes (mean = 40 minutes). The concordance rate between GeneSearch assay and post-operative serial sectioning was 95.6% (215/225), with a sensitivity of 86.1% (31/36), compared with 94.7% (213/225) and 72.2% (26/36)respectively for ⅡC. The size of metastatic foci correlated with the Ct value of CK19 and mammaglobin (P ＜0.01). Conclusions GeneSearch assay for intraoperative detection of metastase in SLN has a satisfactory performance and demonstrates a relatively higher sensitivity than ⅡC. The potential clinical application still requires further evaluation of larger number of cases.     

hMAM在乳腺癌前哨淋巴结中的表达及其临床意义分析

探讨人乳腺球蛋白(human mammary globulin,hMAM)在乳腺癌前哨淋巴结(sentinel lymph node,SLN)中的表达及其临床意义。RT-PCR检测20例乳腺癌组织及正常淋巴结组织中hMAM的表达;SLN定位后通过RT-PCR与常规病理检测SLN转移情况,并进行比较;分析SLN转移与临床病理的关系。乳腺癌组织中有18例过表达,表达率为90.0%,而正常淋巴结中hMAM不表达,两者差异有统计学意义(P<0.01)。常规病理检查转移率为50.98%,RT-PCR法检测的阳性率提高到70.59%,RT-PCR法比常规病理检测的检出率高19.61%,两者之间差异显著(χ2=38.28,P<0.01)。SLN无转移组、微转移组、转移组与肿瘤位置、ER表达及病理类型之间差异均不显著(P>0.05)。hMAM可作为辅助判断SLN是否转移的标志。     

前哨淋巴结病理超分期检测肿瘤微转移在子宫颈癌中的应用

目的探讨前哨淋巴结(sentinel lymph node,SLN)活检和病理超分期技术在子宫颈癌中检测微转移的应用。方法收集子宫颈癌淋巴结组织标本51例行病理超分期检测:组织标本连续切片20张,厚度4μm;第3、8、13、18切片行免疫组化SP法染色检测CK(AE1/AE3),余行HE染色,镜下观察。结果 51例患者SLN经病理超分期检测,5例(9.8%)检出肿瘤转移,其中2例(3.9%)微转移,3例(5.9%)孤立的肿瘤细胞。随访至今尚无肿瘤复发。结论病理超分期技术可以检出常规HE染色可能漏检的肿瘤微转移灶; SLN活检旨在作为盆腔淋巴结切除的可能代替治疗方案,以避免过度治疗、减少术后并发症,提高患者术后生存质量。     

乳腺癌术中前哨淋巴结169枚印片细胞学诊断分析

目的探讨印片细胞学检查在乳腺癌术中前哨淋巴结诊断的价值,提高术中快速诊断的准确率。方法对67例乳腺癌患者的169枚前哨淋巴结同时进行术中冷冻切片检查及印片细胞学检查,与术后石蜡切片诊断对比分析。结果以淋巴结枚数为单位,169例术中前哨淋巴结冷冻切片确诊163例,确诊率96.45%;印片细胞学确诊162例,确诊率95.86%;两者联合诊断,共同确诊166例,确诊率98.22%。结论乳腺癌术中前哨淋巴结印片细胞学检查与冷冻切片检查相结合有互补作用,联合应用可提高术中前哨淋巴结诊断准确率。     

乳腺癌组织中HPV病毒超微结构及DNA检测的研究

目的:探讨广西妇女乳腺癌与HPV感染的关系,并从超微结构水平上观察HPV以及HPV感染后乳腺细胞的形态学改变.方法:使用透射电镜和DNA分子原位杂交两种方法对手术切除的新鲜的40例乳腺癌组织、30例乳腺良性病变组织以及30例乳腺纤维腺瘤旁正常乳腺组织进行HPV16/18检测.结果:电镜下三组HPV的阳性率分别为50%、13.3%、6.7%,差异有统计学意义(P<0.05),细胞核中观察剑的HPV样病毒颗粒,直径约为40nm,有的片状分布,呈假结晶排列,有的聚集成团,无明显结构形成.含有上述HPV样颗粒的细胞,细胞核异形明显.乳腺癌中有淋巴结转移组与无淋巴结转移组的阳性率分别为75%、12.5%,差异也有统计学意义(P<0.05).分子原位杂交技术检测三组的阳性率分别为70%、33.3%、20%,差异有统计学意义(P<0.05).乳腺癌中有淋巴结转移组与无淋巴结转移组的阳性率分别为91.7%、37.5%,差异有统计学意义(P<0.05).电镜下69.2%HPV样病毒颗粒阳性病例中可检测到HPV16/18 DNA存在.结论:广西地区大多数乳腺癌妇女确实存在着HPV16/18感染.检测乳腺癌中HPV16/18 DNA可以作为临床上判断有无淋巴结转移的一个参考指标.该研究对乳腺癌的病因、诊断和预后,降低发病率和死亡率有重要的意义.     

乳腺手术中细胞印片的病理诊断价值

手术中快速病理诊断方法包括冷冻切片、快速石蜡切片和细胞学诊断。由于冷冻切片机设备的不断改进,使冷冻切片质量有了保证,加上具有操作时间短等优点而被广泛应用。快速石蜡切片由于操作步骤较烦琐,时间较长而很少被应用。细胞印片技术操作简单,在更短时间内即可对所送标本做出定性诊断。我们通过对772例术中乳腺标本分别进行细胞印片和冷冻切片检查,并对二者进行比较,探讨细胞印片法在乳腺块中的应用价值。     

吲哚胺2,3-双加氧酶参与乳腺癌患者免疫耐受的研究

目的:研究吲哚胺2,3-双加氧酶(Indoleamine2,3-dioxygenase,IDO)在乳腺癌组织和引流淋巴结中的表达及调节性T细胞在相应组织内的分布,探讨IDO在乳腺癌免疫耐受中的作用机制.方法:收集26例乳腺癌患者的癌组织、癌旁正常乳腺、引流淋巴结和10例乳腺良性病变组织,用RT-PCR法检测IDO mRNA表达,用免疫组织化学法检测IDO和Foxp3蛋白表达.结果:乳腺癌引流淋巴结中IDO mRNA表达水平及IDO表达阳性指数[(19.59±7.65)%]高于原发乳腺癌组织[IDO表达阳性指数(13.16±7.82)%](P<0.05),乳腺癌组织中IDO mRNA表达水平及IDO表达阳性指数高于乳腺良性病变组织[IDO表达阳性指数(3.24±1.30)%]和癌旁正常乳腺组织[IDO阳性细胞指数(2.70±1.53)%](P均<0.05).乳腺癌组织中IDO表达水平与肿瘤临床分期和淋巴结转移相关(P<0.05).乳腺癌引流淋巴结中Foxp3阳性细胞指数[(6.13±2.31)%]高于乳腺原发癌[(3.50±1.04)%],乳腺癌组织中Foxp3阳性细胞指数高于乳腺良性病变[(0.71±0.42)%]和癌旁正常乳腺组织[(0.55±0.34)%](P均<0.05).乳腺癌和引流淋巴结中IDO的表达水平与Treg细胞的分布间均正性相关(r~2=0.449,r~2=0.454,P均<0.05).结论:IDO在乳腺癌细胞中表达增高,并伴随乳腺癌和引流淋巴结中Treg细胞比例升高,提示IDO表达增高有可能通过募集Treg细胞参与肿瘤和引流淋巴结内的免疫耐受.     

乳腺癌术中腋窝前哨淋巴结细胞学印片快速免疫细胞化学检查的应用价值

目的:探索快速免疫细胞化学技术（RICC）应用于乳腺癌术中腋窝前哨淋巴结（SLN）印片检查的可行性及准确性。方法:收集2020—2021年复旦大学附属肿瘤医院乳腺癌患者术中送检的SLN,大体观察SLN转移情况,同时行细胞学印片（TIC）及RICC检测并分别计数染色操作和阅片时长,RICC检测指标为广谱细胞角蛋白（CKp...     

乳腺癌前哨淋巴结微转移分子检测及其临床意义

目的探讨乳腺癌前哨淋巴结(SLN)定位和SLN微转移检测的临床意义。方法对66例乳腺癌患者行术前Y探测仪SLN定位,用RTPCR法检测SLN中CK19mRNA的表达。同时与常规病检法比较其检测敏感性。并比较转移组、微转移组、无转移组患者的临床病理资料。结果SLN定位成功率为97%,RTPCR法与常规病检法转移的检出率相比较差异有统计学意义(P<0.05)。在常规病检阴性的38例淋巴结中,RTPCR法检出8例有微转移。同时乳腺癌转移组与微转移组患者在肿物大小与淋巴管浸润上有相似性,而同无转移组差异有统计学意义(P<0.05)。结论RTPCR法较常规病理检查更为敏感,通过SLN定位和RTPCR的联合使用,可明显提高乳腺癌SLN微转移的检出率。同时也证明RTPCR法是可靠的,SLN微转移有可能作为肿瘤预后的指标。     

细胞涂片检测乳腺癌雌激素受体的价值

应用免疫组化ABC法对30例乳腺癌染色检测雌激素受体的结果表明,针吸细胞涂片或印片阳性率为80%,冰冻切片的阳性率为86.7%,两者符合率93%;细胞涂片与组织切片的免疫组化染色积分呈正相关。(γ=0.77,P<0.01)。结果提示针吸细胞涂片或印片是一种可靠的检测雌激素状况的方法。     

Rapid intraoperative immunohistochemical evaluation of sentinel lymph nodes for metastatic breast carcinoma.

OBJECTIVE: Sentinel lymph node (SLN) biopsy is an integral part of the surgical management of patients with breast cancer. Rapid immunohistochemistry (RIHC) has the potential to increase detection of metastatic carcinoma at the time of frozen section consultation. The authors assessed the accuracy and turnaround time of a newly developed RIHC method for pancytokeratin (RIHC-CK). METHODS: Sixty-six SLNs from 32 patients with breast carcinoma were examined for metastasis using the Zymed Sentinel Lymph Node Rapid IHC Kit. Intraoperative frozen sections (6 mum) of the SLNs were incubated with Zymed anti-pan-cytokeratin/HRP conjugate, diaminobenzidine (DAB), and stained with hematoxylin. Slides were ready within 8 minutes and were interpreted as positive or negative for metastatic carcinoma. Results were compared with previous intraoperative touch preparations, frozen sections, hematoxylin and eosin (Perm H&E), and AEl/3-immunostained permanent sections (Perm CK). RESULTS: Fourteen lymph nodes (19%) in 13 patients tested positive for metastatic carcinoma in Perm H&E, the gold standard. RIHC-CK had the highest sensitivity (92%) of the intraoperative tests, compared with touch preparations (64%) and frozen sections (80%). RIHC-CK showed 94% accuracy, compared with 96% (frozen section) and 93% (touch preparation). The RIHC technique took 8 minutes and was easy to perform and interpret. CONCLUSIONS: Zymed RIHC is a sensitive method for detecting breast cancer metastases in SLNs. The speed, accuracy, and ease of interpretation of the test allow for recognition of micrometastases (<2 mm) that might otherwise be undetectable by current methods of intraoperative evaluation. The prognostic significance and effect on surgical management of micrometastases in SLNs have yet to be determined.     

Evaluation of axillary sentinel lymph node biopsy by immunohistochemistry and multilevel sectioning in patients with breast carcinoma

BACKGROUND: Axillary lymph node dissection for evaluation of the presence or absence of metastatic disease is the single most important prognostic factor for patients with newly diagnosed primary breast cancer. Recently, sentinel lymph node (SLN) biopsy is being investigated as an alternative to the evaluation of the entire axilla. We evaluated whether the application of multilevel sectioning and immunohistochemistry in SLNs will increase the accuracy of detection of metastatic deposits. METHODS: Between October 1998 and July 1999, 38 patients with breast carcinoma (25 ductal, 5 lobular, 4 tubular, and 4 mixed ductal and lobular) underwent successful SLN biopsy followed by complete axillary node dissection. Sentinel lymph nodes were localized with a combination of isosulfan blue dye and radionuclide colloid injection. Frozen sections and permanent sections of SLNs were examined. All negative SLNs were examined for micrometastases by 3 additional hematoxylin-eosin (H&E)-stained sections and immunohistochemistry with the cytokeratins AE1/AE3. RESULTS: Sentinel lymph nodes were successfully identified surgically in 38 (93%) of 41 patients. There was a 97% correlation between the results of the frozen sections and the permanent H&E-stained sections. Twelve (32%) of 38 patients showed evidence of metastatic disease in their SLN by routine H&E staining. In 7 (58%) of 12 patients with positive nodes, the sentinel node was the only positive node. The 26 patients with negative SLN examination by H&E were further analyzed for micrometastases; 5 (19%) were found to have metastatic deposits by immunohistochemistry. Of these patients, 2 were also converted to node positive by detection of micrometastatic disease by examination of the additional H&E levels. CONCLUSIONS: Sentinel lymph nodes can be accurately identified in the axilla of breast cancer patients. Evaluation of SLNs provides reliable information representative of the status of the axilla in these patients. Immunohistochemistry and, to a lesser degree, detailed multilevel sectioning are able to further improve our ability to detect micrometastatic disease in SLNs of breast cancer patients.     

Histopathological work-up and interpretation of sentinel lymph nodes removed for vulvar squamous cell carcinoma

Aims:  To evaluate the work-up of sentinel lymph nodes (SLNs) removed for vulvar pT1–pT2 squamous cell carcinoma (SCC). Inguinal lymphadenectomy yields metastases in only 30% of cases. Patients with missed inguinal disease, however, have a risk of dying from systemic disease. SLN dissections reduce morbidity, but work-up should reliably identify metastatic disease.
Methods and results:  All SLNs removed from 38 patients with pT1–pT2 SCC and clinically negative inguinal lymph nodes were submitted for frozen section analysis. When negative, SLN were formalin-fixed, sectioned entirely at 330-μm intervals to produce three slides per millimetre [two haematoxylin and eosin (H&E) stained slides; one slide for immunohistochemistry]. If screening of H&E-stained sections was negative, all remaining slides were subjected to immunohistochemistry with an antibody to cytokeratin. Twenty-five of 38 patients (66%) were pN0, 7/38 (18%) had metastases on frozen sections/H&E stains. Immunohistochemistry detected micrometastases in two patients and single tumour cells and anucleate cell structures in four patients. In 12/13 patients the SLN metastases, including all single-cell deposits, were from lichen sclerosus (LS)-associated SCC. Twelve of 13 patients with metastases had a pT2 SCC.
Conclusions:  Micrometastases and single tumour cell deposits in SLNs are typical of LS-associated vulvar SCC. Single tumour cell deposits in SNLs should be regarded as 'positive'. Identification requires serial sectioning and immunohistochemical analysis of all removed SLNs.     

Importance of assessing CK19 immunostaining in core biopsies in patients subjected to sentinel node study by OSNA

Analysis of sentinel lymph node (SLN) by means of One-Step Nucleic Acid Amplification (OSNA) is being used increasingly as a very sensitive and quick method for intraoperative axillary staging in patients with breast cancer. This molecular diagnostic assay detects the expression level of cytokeratin 19 (CK19), a luminal epithelial cell marker broadly expressed in most breast carcinomas and not normally found in lymph nodes. Almost all breast cancers express this cytoskeleton protein, but some breast tumors have been found to lose the expression of CK19. CK19 immunostaining in core biopsies has been recommended in selecting patients eligible for OSNA analysis because SLNs with metastatic involvement by CK19-negative breast cancers may result in a false negative result by OSNA. However, the real frequency of CK19-negative breast cancer has to be elucidated. In this study, we have assessed the frequency and molecular profile of CK19-negative breast carcinomas in three series of cases. The first is a prospective series of 197 breast carcinomas, 111 of which were subjected to SLN evaluation by OSNA. The second is a retrospective series of 41 triple-negative (TN) breast carcinomas, and the third is a retrospective series of 68 breast cancer patients (matched core biopsies and metastatic lymph nodes) that had been evaluated by conventional procedures before the OSNA methodology was adopted in our institution. Our results not only demonstrate that lack of expression of CK19 is infrequent in breast cancers but also that performing CK19 immunohistochemical staining is important on diagnostic core biopsies in taking the decision of using OSNA methodology in the evaluation of sentinel nodes in breast cancer patients.     

Rapid immunohistochemistry of sentinel lymph nodes for metastatic melanoma

Sentinel lymph node (SLN) biopsy is performed on patients with malignant melanoma (MM) to assess the need for selective complete lymphadenectomy. Melanoma metastasis to regional lymph nodes is an important prognostic indicator in patients with MM. This study assesses the sensitivity and specificity of rapid immunohistochemistry (RIHC) in intraoperative delineation of melanoma metastasis to SLN. RIHC for S-100 protein, HMB45, and a melanoma marker cocktail (melan A, HMB45, and tyrosinase) was performed on 71 SLNs obtained from 28 patients with MM. Frozen sections (6 micro thick) on plus slides were fixed for 2 to 3 minutes in cold acetone and then stored at -70 degrees C. The EnVision kit (Dako, Carpinteria, CA) for rapid immunohistochemistry (RIHC) on frozen tissue sections was used, and the staining technique took 19 minutes. Together with preparation of the frozen sections and fixation in acetone, immunostained slides were available in approximately 25 minutes. Of the 71 SNLs examined, 7 showed melanoma metastasis in permanent sections. RIHC of frozen sections detected metastatic melanoma in 6 SLNs, with a sensitivity of 86% for HMB45 and 71% for S-100 protein and the melanoma cocktail and a specificity of 97% for HMB45 and 100% for S-100 and the melanoma cocktail. We conclude that RIHC for HMB45, S-100 protein, and the melanoma cocktail may help detect melanoma metastasis in SLN intraoperatively, leading to total lymph node dissection and obviating the need for 2 surgical procedures. Section folds and background stain can make interpretation difficult. Intraoperative time constraints require a more rapid technique. A recent consensus group has discouraged frozen-section examination of SLN.     

Sentinel lymph node investigation in melanoma: detailed analysis of the yield from step sectioning and immunohistochemistry

AIMS: To evaluate in detail the extent to which step sectioning and immunohistochemical examination of sentinel lymph nodes (SLNs) in patients with melanoma reveal additional node positive patients, to arrive at a sensitive yet workable protocol for histopathological SLN examination. METHODS: The study comprised 29 patients with one or more positive SLN after a successful SLN procedure for clinical stage I/II melanoma. SLNs were lamellated into pieces of approximately 0.5 cm in size. One initial haematoxylin and eosin (H&E) stained central cross section was made for each block. When negative, four step ribbons were cut at intervals of 250 microm. One section from each ribbon was stained with H&E, and one was used for immunohistochemistry (IHC). RESULTS: When taking the cumulative total of detected metastases at level 5 as 100%, the percentage of SLN positive patients increased from 79%, 83%, 83%, 90% to 93% in the H&E sections through levels 1-5, and with IHC these values were 83%, 86%, 90%, 97%, and 100%, respectively. One of six patients in whom metastases were detected at levels 2-5 only had metastases in the subsequent additional lymph node dissection. CONCLUSIONS: Multiple level sectioning of SLNs (five levels at 250 microm intervals) and the use of IHC detects additional metastases up to the last level in melanoma SLNs. Although more levels of sectioning might increase the yield even further, this protocol ensures a reasonable workload for the pathologist with an acceptable sensitivity when compared with the published literature.     

Histopathological patterns of melanoma metastases in sentinel lymph nodes

AIMS: Sentinel lymph node biopsy (SLNB) is an important component in the staging and treatment of cutaneous melanoma (CM). The medical literature provides only limited information regarding melanoma sentinel lymph node (SLN) histology. This report details the specific histological patterns of melanoma metastases in sentinel lymph nodes (SLNs) and highlights some key factors in evaluating SLNs for melanoma. METHODS: From 281 SLNB cases between June 1998 and May 2002, 79 consecutive cases of SLN biopsies positive for metastases from CM were retrospectively reviewed. The important characteristics of the SLNs and the metastatic foci are described. RESULTS: The median size of positive SLNs was 17 mm (range, 5-38). SLNs had a median of two metastatic foci (range, 1-11), with the largest foci being a median of 1.1 mm in size (range, 0.05-24). S-100 and HMB-45 staining was positive in 100% and 92% of the detected metastatic foci, respectively. The metastatic melanoma cells were epithelioid, spindled, and mixed in 86%, 5%, and 9% of cases. Metastatic foci were most often (86%) found in the subcapsular region of the SLN. Benign naevic cells were found coexisting in 14% of positive SLNs. CONCLUSIONS: Staining for S100 is more sensitive than HMB-45 (100% v 92%), but HMB-45 staining helped to distinguish benign naevic cells from melanoma. The subcapsular region was crucial in SLN evaluation, because it contained the metastases in 86% of cases. Evaluation of the subcapsular space should not be compromised by cautery artefacts or incomplete excision of the SLN.     

Intraoperative sentinel lymph node evaluation in patients with node-positive breast cancer status post neoadjuvant systemic therapy - An institutional experience

Recent studies have shown the feasibility and utility of sentinel lymph node (SLN) biopsy in patients with biopsy proven node-positive breast cancer after neoadjuvant chemotherapy. We reviewed our experience in intraoperative SLN evaluation in such cases and its effect on axillary management. A retrospective analysis of breast cancer patients (2015–2018) with a biopsy-proven positive axillary lymph node, who received neoadjuvant systemic therapy and underwent intraoperative SLN assessment was performed. Intraoperative SLN assessment results were compared with final pathology. Its accuracy and effect on axillary management is summarized. We identified 106 patients with positive axillary lymph node and neoadjuvant systemic therapy between the ages of 28 and 75 years who had SLN biopsy and lumpectomy (33) or mastectomy (73). Three or more SLNs were identified in 91 cases (86 %). The previously biopsied lymph node was identified as one of the sentinel lymph nodes in 93 cases (88 %). There is a high concordance rate between frozen section diagnosis and final diagnosis on sentinel lymph nodes. No false positive case and seven false negative frozen section diagnosis cases (diagnosed as negative on frozen section and positive on permanent sections) were identified. False-negative frozen section diagnosis correlated with low-volume nodal disease and obscuring tumor bed changes. Almost half of the positive lymph nodes were converted to negative after neoadjuvant chemotherapy. SLN biopsy with intraoperative frozen section evaluation after neoadjuvant systemic therapy in node-positive patients is an effective way to minimize axillary surgery.     

Metastatic melanoma volume in sentinel nodes: objective stereology-based measurement predicts disease recurrence and survival

Aims:  Sentinel lymph node (SLN) status is the most important prognostic factor in intermediate thickness melanoma. The amount of metastatic disease in positive SLNs varies greatly between patients, and this tumour burden appears to influence the prognosis of node-positive patients. The aim was to use objective stereological techniques to correlate accurately total SLN tumour burden with recurrence and patient survival.
Methods and results:  SLNs from 327 patients were examined by complete step sectioning and immunohistochemistry. The total metastasis volume (TMV) of 156 positive SLNs from 99 patients (30.3%) was measured using stereological methods based on the 2D-nucleator and Cavalieri's principle. The maximum metastasis diameter was also measured. These two measurements were correlated with disease recurrence and patient survival. The mean TMV for SLN+ patients was 10.5 mm³ (median 0.05 mm³; range 0.0001–623.7 mm³). Median follow-up was 26.3 months. On multivariate analysis, TMV was an independent predictor of recurrence when corrected for primary tumour thickness ( P  = 0.001) and was a stronger prognosticator compared with the maximum metastasis diameter ( P  < 0.0001 versus P  = 0.01).
Conclusions:  Combining total step sectioning of SLNs with stereological assessment of metastases, we found metastasis volume to be a highly significant predictor of disease recurrence and survival.     

One-step nucleic acid amplification (OSNA) for intraoperative evaluation of sentinel lymph node status in breast cancer: a comparative study between CK19 protein expression and CK19 mRNA level in primary tumors and lymph node metastasis

The one-step nucleic acid amplification (OSNA) method is an increasingly used procedure for intraoperative analysis of sentinel lymph node (SLN) status in breast cancer patients. It measures cytokeratin19 (CK19) mRNA copy numbers in homogenized samples of SLN; CK19 has been chosen for identifying node metastasis because most breast cancers express this molecule. However, to avoid false-negative OSNA results, testing the preoperative needle core biopsy (NCB) of breast carcinomas for CK19 by immunohistochemistry (IHC) has been recommended. This procedure relies on the assumption that protein expression is strictly related to mRNA expression. We developed this study to evaluate if IHC gives similar result to the molecular assay. In a series of breast cancer patients with axillary metastasis, corresponding surgically resected tumor and metastatic lymph node specimens have been tested for CK19 protein by IHC and for CK19 mRNA by real-time PCR; furthermore, CK19 immunostaining has been performed in NCBs when available. Statistical analysis revealed that (1) the immunohistochemical evaluation of CK19 in NCB is a reliable test, reflecting protein expression in the whole tumor and in the metastatic lymph node; (2) there is no correlation between CK19 protein expression and CK19 RNA level neither within the primary breast cancer nor within the metastatic node; moreover, no correlation as well has been found between protein expression in NCB and mRNA level in metastatic lymph nodes. Thus, our results suggest that there is no evidence-based reason to stain every NCB for CK19 before performing OSNA in patients with breast cancer.