Journal of Laboratory Physicians最新文献

Background Lupus nephrtis in children is associated with high morbidity and mortality. The incidence of childhood systemic lupus erythematosus (SLE) ranges from 3.3 to 8.8/100000 children with a higher Asian preponderance. The predominance of SLE in female pediatric patients increases gradually with age to the values observed in adults. Objectives  To assess the clinical, immunological, and histopathological spectrum of childhood lupus nephritis in northeast India and explore the relationship between clinical, biochemical, serological, and histopathological findings. Materials and Methods  A retrospective descriptive study was performed over 8 years. Histopathology slides were reviewed by two pathologists, whereas other details were collected from patients' records. Statistical Analysis  Statistical analysis was based on the chi-square test and a p -value < 0.05 was considered statistically significant. Results  Fifty-three cases of lupus nephritis were included in the study. The patients' age ranged from 5 to 18 years with a mean age of 14.5 years and a female: male ratio of 6.5:1. Edema and hypertension were the commonest clinical presentations, whereas proteinuria was the commonest presenting laboratory parameter. Amongst all the immunological markers, dsDNA was the commonest. Histopathologically, predominantly study population belonged to class IV lupus nephritis. The patients with class IV showed a statistically significant correlation with proteinuria and hematuria at the time of diagnosis. Immunological markers, namely, ANA and anti-ds-DNA positivity were significantly associated with advanced renal histopathology. Conclusion  cSLE in northeast India presents mostly as Class IV LN presenting mostly with deranged laboratory parameters and preponderance of various immunological markers and clinical presentations.

Merkel cell carcinoma (MCC) is a rare primary neuroendocrine tumor of the skin. It has an aggressive biological behavior and shows early local and distant metastasis. Diagnosis of MCC is a challenge and requires confirmation by immunohistochemistry (IHC). However, metastasis of MCC to the stomach is extremely uncommon and is rarely reported in the literature. We hereby describe a patient with gastric metastasis of MCC, who presented with black tarry stool and was finally diagnosed on the basis of clinical history, histology, and IHC.

Introduction  Cesarean scar pregnancy (CSP) is an increasing clinical condition that causes serious maternal morbidity and mortality. This study aimed to evaluate if inflammation markers measured by hemogram can aid in the diagnosis of CSP. Materials and Methods  A total of 86 patients were included in the study. The cases were divided as CSP ( n : 42) and normal pregnancy (NP) ( n : 44). At the time of admission, peripheral blood neutrophils, lymphocytes, monocytes, thrombocytes, systemic inflammatory index (SII) (neutrophil × platelet/lymphocyte), neutrophil-lymphocyte ratio, monocyte-lymphocyte ratio, and platelet-lymphocyte ratio were all measured. CSP and NP diagnoses were made by transabdominal or vaginal ultrasonography. Results  In the CSP group, mean age ( p  < 0.001), gravida ( p  < 0.001), parity ( p  < 0.001), number of surviving children ( p  < 0.001), number of abortions ( p  < 0.001), cesarean number ( p  < .001), dilatation and curettage count ( p  = 0.013), monocyte (M) value ( p  = 0.039) and monocyte/lymphocyte value (MLR) ( p  = 0.035) were significantly higher than the control group. The optimal M value cut-off value was found to be > 0.40, the sensitivity value was 78.57, and the specificity value was 50.00. AUC = 0.632 (SE = 0.061) for the MLR value. The optimal MLR cut-off value was found to be > 0.232, the sensitivity value was 61.90, and the specificity value was 63.64. Conclusion  Hemogram parameters, which are simple, inexpensive, and easily accessible, M and MLR are significantly higher in the diagnosis of CSP and can be used as an auxiliary parameter for ultrasonography.

Introduction  Cancer stem cell markers are now being tried in various cancers as prognostic markers including GI cancer but these kinds of studies are sparse in Indian population. Materials and Methods  This study conducted over a period 50 months. Hematoxylin and eosin-stained slides were screened for grading of the tumor, extent of invasion of tumor, confirmation of metastasis, and staging was done. Immunohistochemical expression of CD44 was graded on the basis of percentage of tumor cells positive for staining. Statistical analysis was done and results were tabulated. Results : A total of 40 cases of GI cancer were studied. Ascending colon (37.5%) was the common site involved, 37 cases (92.5%) showed invasion beyond the muscularis externa. Most tumors were poorly differentiated (37.5%). Also, 50% of lymph nodes showed tumor deposits. The majority of the cases were in stage II (40%). There was a significant correlation between histopathological type of differentiation with lymph node metastasis and staging of tumor, lymph node metastasis also had significant association with staging. Grade 2, CD 44 expression was most common followed by Grade 3. Significant association was observed between histopathological differentiations of tumor with CD44 expression. Tumors that are invading beyond muscularis externa and lymph node-positive cases showed moderate to high CD44 expression. Conclusion  CD44 expression was significantly noted in poorly differentiated tumors. Increased expression was also noted in cases of tumors invading beyond muscularis externa and lymph node metastasis. Combination of CSC markers will increase the sensitivity and specificity and predict better overall survival in GI tumors.

Background  Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created high demand for molecular kits and consumables for mass screening of suspected individuals. Direct real-time polymerase chain reaction (RT-PCR) assay without nucleic acid extraction has several advantages in saving testing time and cost and helps in the rapid reporting of SARS-CoV-2. The present study evaluated the analytical performance of four SARS-CoV-2 RT-PCR for direct RT-PCR testing using preheated specimens. Methods  A total of 100 clinical specimens were selected and divided into three different groups: (1) group I: 20 SARS-CoV-2 positive specimens with high viral load, viz., low Ct values (< 30 Ct), (2) group II: 50 SARS-CoV-2 positive specimens with low viral load, viz., high Ct values (> 30 Ct), and (3) group III: 30 SARS-CoV-2 negative specimens. Specimens were heat-inactivated at 70°C for 10 minutes and cooled down at 4°C and were evaluated for standard and direct RT-PCR method by using ViralDtect-II Multiplex Real-Time PCR kit, TaqPath COVID-19 Combo kit, COVIDsure Pro Multiplex RT-PCR kit, and Hi-PCR Coronavirus (COVID-19) Multiplex Probe PCR kit. Results  Results showed that except ViralDtect-II kit, the other three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit were able to amplify all the SARS-CoV-2 genes in the direct RT-PCR method using preheated specimens. In group I specimens, 100% sensitivity was observed in all three RT-PCR kits. In group II specimens, COVIDsure Pro kit was found to be superior among other kits. Conclusion  Direct RT-PCR method during pandemic situation is valuable and cost effective for the detection of SARS-CoV-2. All three TaqPath COVID-19 Combo kit, COVIDsure Pro kit, and Hi-PCR Coronavirus (COVID-19) RT-PCR kit can be used for direct RT-PCR method and COVIDsure Pro kit performance was found to be superior among all.

Objective  Microbiological confirmation of tuberculosis (TB) in pediatric cases is challenging due to its paucibacillary nature and difficulty in specimen collection. This study aimed to validate stool as an alternative sample for the diagnosis of pediatric pulmonary TB via Xpert MTB/RIF (Xpert) assay. Materials and Methods  This cross-sectional study included 75 pediatric patients up to 10 years of age with signs and symptoms suggestive of TB. From each recruited patient, pulmonary and stool samples were collected in a sterile container. The collected samples were subjected to Ziehl-Neelsen staining, BACTEC MGIT 960 culture (MGIT), Xpert, and in-house multiplex polymerase chain reaction for TB diagnosis. Results  About 13.33% (10/75) of the pulmonary samples and, of them, 50% (5/75) of the stool samples were positive by Xpert assay. The sensitivity and specificity of Xpert assay with stool and pulmonary samples were 50 (95% confidence interval [CI]: 18.71-81.29%) and 100% (95% CI: 94.48-100%), respectively. Conclusion  The Xpert assay on stool samples showed limited sensitivity and good specificity in the diagnosis of pulmonary TB. Therefore, it can be proposed as an alternative screening sample to diagnose TB in pediatric cases for which getting a respiratory sample is extremely difficult. However, further studies with greater number of samples and multiple baseline variables are required to support our findings. Strategies to optimize stool Xpert assay should be performed to enhance the sensitivity of this method to detect Mycobacterium tuberculosis in children.

Introduction  Vaccination has shown to be protective against severe coronavirus disease 2019 by various studies. However, the vaccine efficacy was demonstrated to be less against the emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Both vaccine- and infection-induced immunity against SARS-CoV-2 may prevent reinfection and severity. Our study aims to assess and compare the humoral response in heterogeneous population based on infection and vaccination status along with hybrid immunity. Methods  A retrospective, observational study of 2,545 adults was conducted. The study groups comprised of group I ( n  = 309) naive with a single dose of vaccination, group II ( n  = 357) infected and unvaccinated, group III ( n  = 590) completely vaccinated with two doses of vaccine, group IV ( n  = 70) booster dose, group V ( n  = 602) with hybrid immunity (pre-vaccination infection), and group VI ( n  = 617) with breakthrough infection (post-vaccination infection). Data pertaining to demographic details, clinical presentations, reverse transcription-polymerase chain reaction, anti-SARS-CoV-2 total antibodies immunoglobulin G (IgG), neutralizing antibodies by anti SARS-CoV-2 sVNT (surrogate virus neutralization test), S1/S2IgG, S-RBD (receptor-binding domain), and ChAdOx1-nCov-19 (Covishield) vaccination were retrieved from electronic health records. Results  The mean levels of neutralizing antibodies of group V were S1/S2, RBD (10.5/14.3 times), and sVNT (84.44%) and group VI had S1/S2, RBD (11.4/11.8 times), and sVNT (78.07%) when compared to group III. We also observed a statistically significant higher immune response in group V and VI than group I and II. A higher percentage (18.2%) of group II individuals had severe disease when compared to group V and VI (6.5/10.8%). Conclusion  A single dose of ChAdOx1 vaccine gives robust antibody responses in previously infected individuals and may confer long-term hybrid immunity following booster vaccination.

Objectives  Methotrexate (MTX) has anticancer therapeutic potential with multiple doses-related adverse effects and toxicities. Immunoassays for therapeutic monitoring of serum MTX have their own limitations. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered as the reference method; however, commercially availability of them is limited. We aimed to adapt/develop an in-house LC-MS/MS method for therapeutic monitoring of serum MTX. Materials and Methods  Serum protein precipitation was performed using acetonitrile-water containing 250 μM solution of aminoacetophenone as internal standard (IS). Chromatographic separation was achieved on a C18 column with mobile phase of 0.1% solution of formic acid (solvent A) and acetonitrile (solvent B) at a flow rate of 0.4 mL/min. MS was performed under positive ion mode with mass transition for MTX and IS as m/z 455.1→308.1 and 136.2→94.1, respectively. The method was validated by following Bioanalytical Method Validation Guidance for Industry, 2018 and applied on leukemia patients' samples on MTX therapy. Results  The correlation coefficient of eight serially diluted calibration standards of 0.09 to 12.5 μM was >0.99 and had linearity with > 95% precision and accuracy at analytical quality control levels. The lower limit of MTX quantification achieved was 0.09 μM with good intensity and sharp peak as compared with blank sample. The total run time of the assay was 5 minutes. The serum MTX levels obtained by this method in leukemia patients exhibited clinical correlation and an excellent agreement with commercial immunoassay used in parallel. Conclusion  We were able to develop a rapid, sensitive, and cost-effective LC-MS/MS method suitable for therapeutic drug monitoring of MTX in routine clinical diagnostic laboratories.

Background  Breast cancer is an epithelial malignancy; however, stroma plays a key role with its stimulatory and inhibitory factors in modulating tumor invasion and metastasis. CD10, a matrix metalloproteinase, is known to regulate cell adhesion, migration and helps in determining the progression of tumor. This knowledge helps to identify specific signals that promote growth, dedifferentiation, invasion, metastasis and serve as target for better therapeutic management. Objectives  The aim of this study was to estimate frequency of expression of stromal CD10 and assess its prognostic significance in breast carcinomas by correlating with known prognostic factors. Materials and Methods  Morphological parameters of 62 cases of carcinoma breast were studied on H&E (hematoxylin and eosin) stained sections and expressions of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2/neu), and CD10 on manually constructed tissue microarray sections by immunohistochemistry (IHC). Staining pattern, percentage of stained cells, and intensity of stains were evaluated and IHC scoring of all markers was done. CD10 scores were correlated with the known prognostic factors (ER, PR, and HER2/neu). A p -value less than 0.05 was considered as significant. Results  Stromal expression of CD10 was found in 82.3% of cases and it was significantly associated with increasing tumor size ( p  = 0.012), increasing tumor grade ( p  = 0.001), lymph node metastasis ( p  = 0.018), necrosis ( p  = 0.008), lymphovascular invasion ( p  = 0.008), ER negativity ( p  = 0.001), PR negativity( p  = 0.007), HER 2 positivity ( p  = 0.012), triple-negative molecular subtypes ( p  = 0.001), and poor prognostic groups ( p  = 0.01). Conclusion  CD10 can be used as an independent prognostic stromal marker and this will help to envisage new therapeutic strategies.