Culture Drug Sensitivity Test Research Articles

Development and clinical validation of a dual ddPCR assay for detecting carbapenem-resistant Acinetobacter baumannii in bloodstream infections.

Acinetobacter baumannii (A. baumannii, AB) represents a major species of Gram-negative bacteria involved in bloodstream infections (BSIs) and shows a high capability of developing antibiotic resistance. Especially, carbapenem-resistant Acinetobacter baumannii (CRAB) becomes more and more prevalent in BSIs. Hence, a rapid and sensitive CRAB detection method is of urgent need to reduce the morbidity and mortality due to CRAB-associated BSIs. A dual droplet digital PCR (ddPCR) reaction system was designed for detecting the antibiotic resistance gene OXA-23 and AB-specific gene gltA. Then, the specificity of the primers and probes, limit of detection (LOD), linear range, and accuracy of the assay were evaluated. Furthermore, the established assay approach was validated on 37 clinical isolates and compared with blood culture and drug sensitivity tests. The dual ddPCR method established in this study demonstrated strong primer and probe specificity, distinguishing CRAB among 21 common clinical pathogens. The method showed excellent precision (3 × 10-4 ng/μL, CV < 25%) and linearity (OXA-23: y = 1.4558x + 4.0981, R2 = 0.9976; gltA: y = 1.2716x + 3.6092, R2 = 0.9949). While the dual qPCR LOD is 3 × 10-3 ng/μL, the dual ddPCR's LOD stands at 3 × 10-4 ng/μL, indicating a higher sensitivity in the latter. When applied to detect 35 patients with BSIs of AB, the results were consistent with clinical blood culture identification and drug sensitivity tests. The dual ddPCR detection method for OXA-23 and gltA developed in this study exhibits good specificity, excellent linearity, and a higher LOD than qPCR. It demonstrates reproducibility even for minute samples, making it suitable for rapid diagnosis and precision treatment of CRAB in BSIs.

Retracted: Correlation between Sputum Bacterial Culture Positive Rate and Drug Sensitivity Test Results and Disease Severity in Inpatients and Its Clinical Significance: A Systematic Review and Meta-Analysis.

[This retracts the article DOI: 10.1155/2022/5102100.].

Impact of molecular diagnostic tests on diagnostic and treatment delays in tuberculosis: a systematic review and meta-analysis

BackgroundCountries with high TB burden have expanded access to molecular diagnostic tests. However, their impact on reducing delays in TB diagnosis and treatment has not been assessed. Our primary aim was to summarize the quantitative evidence on the impact of nucleic acid amplification tests (NAAT) on diagnostic and treatment delays compared to that of the standard of care for drug-sensitive and drug-resistant tuberculosis (DS-TB and DR-TB).MethodsWe searched MEDLINE, EMBASE, Web of Science, and the Global Health databases (from their inception to October 12, 2020) and extracted time delay data for each test. We then analysed the diagnostic and treatment initiation delay separately for DS-TB and DR-TB by comparing smear vs Xpert for DS-TB and culture drug sensitivity testing (DST) vs line probe assay (LPA) for DR-TB. We conducted random effects meta-analyses of differences of the medians to quantify the difference in diagnostic and treatment initiation delay, and we investigated heterogeneity in effect estimates based on the period the test was used in, empiric treatment rate, HIV prevalence, healthcare level, and study design. We also evaluated methodological differences in assessing time delays.ResultsA total of 45 studies were included in this review (DS = 26; DR = 20). We found considerable heterogeneity in the definition and reporting of time delays across the studies. For DS-TB, the use of Xpert reduced diagnostic delay by 1.79 days (95% CI − 0.27 to 3.85) and treatment initiation delay by 2.55 days (95% CI 0.54–4.56) in comparison to sputum microscopy. For DR-TB, use of LPAs reduced diagnostic delay by 40.09 days (95% CI 26.82–53.37) and treatment initiation delay by 45.32 days (95% CI 30.27–60.37) in comparison to any culture DST methods.ConclusionsOur findings indicate that the use of World Health Organization recommended diagnostics for TB reduced delays in diagnosing and initiating TB treatment. Future studies evaluating performance and impact of diagnostics should consider reporting time delay estimates based on the standardized reporting framework.

A PDX model combined with CD-DST assay to evaluate the antitumor properties of KRpep-2d and oxaliplatin in KRAS (G12D) mutant colorectal cancer

Patient-derived xenograft (PDX) models are more faithful in maintaining the characteristics of human tumors than cell lines and are widely used in drug development, although they have some disadvantages, including their relative low success rate, long turn-around time, and high costs. The collagen gel droplet embedded culture drug sensitivity test (CD-DST) has been used as an in-vitro drug sensitivity test for patients with cancer because of its high success rate of primary cell culture, high sensitivity, and good clinical relevance, but it is based on an in-vitro cell culture and may not simulate the tumor microenvironment accurately. This study aims to combine a PDX model with CD-DST to evaluate the efficiency of antitumor agents. KRpep-2d, a small peptide targeting KRAS (G12D), and oxaliplatin were used to verify the feasibility of this approach. Whole-exome sequencing and Sanger sequencing were first applied to test and validate the KRAS mutation status of a panel of colorectal cancer PDX tissues. One PDX model was verified to carry KRAS (G12D) mutation and was used for in-vivo and the CD-DST drug tests. We then established the PDX mouse model from the patient with the KRAS (G12D) mutation and obtained viable cancer cells derived from the same PDX model. Next, the antitumor abilities of KRpep-2d and oxaliplatin were estimated in the PDX model and the CD-DST. We found that KRpep-2d showed no significant antitumor effect on the xenograft model or on cancer cells derived from the same PDX model. In contrast, oxaliplatin showed significant inhibitory effects in both tests. In conclusion, the PDX model in combination with the CD-DST assay is a comprehensive and feasible method of evaluating the antitumor properties of compounds and could be applied for new drug discovery.

Role of Collagen Gel Droplet-Embedded Culture-Drug Sensitivity Testing (CD-DST) for Assessing the Sensitivity of Gastric Cancer to Chemotherapy Drugs Combined with Other Cancer Therapeutic Drugs.

Chemosensitivity tests have long been a widely discussed research topic. Our group performed collagen gel droplet-embedded culture-drug sensitivity testing (CD-DST) of patients with advanced gastric cancer during the period from December 2012 to December 2017. To verify how CD-DST should be used, we invested correlations of sensitivities to cisplatin (CDDP), docetaxel (DOC), paclitaxel (PTX), and CPT11 with clinical outcome. Patients with advanced gastric cancer underwent gastrectomy with lymph node dissection at Nippon Medical School Tama Nagayama Hospital, and surgical samples were retrospectively examined by CD-DST to assess chemosensitivity. The patients later received adjuvant chemotherapy as standard adjuvant therapy or chemotherapy. The CD-DST test was not performed for S-1 because it is commonly used in chemotherapy for gastric cancer. Although oxaliplatin has also recently become a key drug for advanced gastric cancer, it had not been adopted for gastric cancer in 2012, so CD-DST testing was not performed. The χ2 test was used for all statistical analyses. A p-value of <0.05 was assumed to indicate statistical significance. Three-year survival rates were estimated using the Kaplan-Meier method, and the log-rank test was used to compare the obtained curves. Of the tumors from gastric cancer patients, 67.0% (77/115) could be cultured. The rate of sensitivity was 41.1% (30/73) for CDDP, 82.6% (57/69) for DOC, 82.8% (58/70) for PTX, and 49.2% (33/67) for CPT11. CDDP sensitivity and outcome were not correlated in patients who received CDDP. Sensitivities to CDDP, DOC, PTX, and CPT11 were not correlated with any patient characteristic. Patients with poorly differentiated adenocarcinoma tended to be sensitive to CDDP (P=0.051). No difference between CDDP sensitivity or outcome was observed in patients receiving CDDP. The CD-DST showed a high sensitivity to DOC and PTX in the present patients.

Correlation between Sputum Bacterial Culture Positive Rate and Drug Sensitivity Test Results and Disease Severity inInpatients and Its Clinical Significance: A SystematicReview and Meta-Analysis.

Objective To systematically evaluate the correlation between the positive rate of sputum bacterial culture and the results of drug sensitivity test and the severity of the disease and its clinical significance, so as to provide evidence-based medicine for clinical application. Methods PubMed, Embase, ScienceDirect, Cochrane Library, China Knowledge Network Database (CNKI), China VIP Database, Wanfang Database, and China Biomedical Literature Database (CBM) online database were used. The retrieval time limit was from the establishment of the database to the present. Data for all included studies were extracted by two independent researchers, and the risk of bias for the quality of each included study was assessed by the Cochrane Handbook 5.1.0 criteria. RevMan5.4 statistical software was used to analyze the collected data by meta. Results In the end, 6 RCT articles were included. Overall, 613 samples were included in 6 RCT studies. The correlation between the positive rate of sputum bacterial culture in inpatients and the severity of the disease was meta-analyzed. The heterogeneity test results showed that Chi2 = 177.20, df = 3, P < 0.00001, and I2 = 98%, indicating that there was obvious heterogeneity among the included research data. It was considered that there was a correlation between the positive rate of sputum bacterial culture and the severity of the disease. The correlation between the results of the drug sensitivity test of inpatients and the severity of the disease was evaluated. The results of the heterogeneity test showed that Chi2 = 0.00, df = 1, P = 1 > 0.05, and I2 = 0%, indicating that there was no heterogeneity among the included research data. In addition, the combined effect of WMD was analyzed by the fixed effect model. The combined effect dose WMD test was Z = 6.58 (P < 0.00001). It was considered that there was a correlation between the results of the drug sensitivity test and the severity of the disease. Conclusion There is a correlation between positive sputum culture and drug sensitivity test results and the severity of the disease in hospitalized patients. In clinical practice, for hospitalized patients, the positive sputum bacterial culture rate and drug sensitivity test results can be used to guide the appropriate use of antibiotics. Due to the low input from the literature, more studies with higher methodological quality and longer follow-up are needed for further validation.

Etiological analysis of infectious keratitis in children from 2007 to 2016

Objective: To analyze the etiological changes of children with infectious keratitis. Methods: Retrospective study. Data of patients diagnosed as bacterial, fungal, and amoebic keratitis from 2007 to 2016, aged no more than 14 years old, were collected in the Department of Ophthalmology, Beijing Tongren Hospital. A total of 649 samples were obtained for routine laboratory culture identification and drug sensitivity tests. There were 361 males and 278 females, aged (5.6±4.4) years. The data were analyzed according to age ≤3 years, 4 to 7 years and>7 years. The qualitative data were analyzed by the Chi-square test. Results: Among 649 samples, 140 were culture positive, and the positive rate was 21.6%. Bacteria were the main pathogens, accounting for 81.4%. The positive rate was 31.0% among bacterial samples (114/368), and the bacterial flora was mainly Gram-positive cocci, followed by Gram-negative bacilli. Streptococcus (34.2%) ranked first, followed by Staphylococcus (27.2%) and Pseudomonas (7.9%). For children no more than 7 years old, Streptococcus pneumoniae was the most common bacterial species, while Staphylococcus epidermidis was most common in those aged 8-14 years. Gram-positive cocci were sensitive to vancomycin. Most bacteria were more sensitive to fluoroquinolones and less sensitive to cephalosporins and aminoguanidine. The detection rate of methicillin-resistant Staphylococcus was 68% (17/25). Gatifloxacin had the highest sensitivity, while the difference between gatifloxacin and moxifloxacin, levofloxacin, ofloxacin were not statistically significant (χ²=0.836,0.358, 1.490; P=0.361,0.550,0.222). A total of 254 specimens were cultured for fungi, of which 22 were positive, and the positive rate was 8.7%. The isolated fungi included Fusarium (34.8%, 8/22), Aspergillus (26.1%, 6/22) and Candida (13.0%, 3/22). The positive rate of fungal culture was 9.2% (11/119) from 2007 to 2011, and 8.1% (11/135) from 2012 to 2016, no significant difference was found between two period (χ²=0.096, P=0.757). Fusarium showed a relatively high sensitivity to terbinafine, but it was not sensitive to fluconazole and itraconazole. The sensitivities of Aspergillus to terbinafine and voriconazole were high, followed by amphotericin. Candida had high sensitivities to amphotericin, fluconazole, itraconazole and voriconazole. In 27 specimens for Acanthamoeba culture, 4 specimens were positive, and the positive rate was 14.8%. Risk factors of Acanthamoeba infection included wearing orthokeratology lenses and trauma. Conclusions: Bacteria are the main pathogenic agent of infectious keratitis in children. Streptococcus pneumoniae is the most common in children aged 7 and below, and Staphylococcus epidermidis is the most common in children aged 8-14. Fungal infection was significantly lower than that of bacteria, mainly Fusarium, Aspergillus and Candida.

Effect of oxaliplatin-based chemotherapy on chemosensitivity in patients with peritoneal metastasis from colorectal cancer treated with cytoreductive surgery and hyperthermic intraperitoneal chemotherapy: proof-of-concept study

BackgroundChemosensitivity testing, including collagen gel droplet‐embedded culture drug sensitivity test, has proven to be a useful tool in therapeutic decision‐making. This retrospective analysis investigated chemosensitivity testing of peritoneal metastases collected during cytoreductive surgery (CRS), and its impact on survival in patients with colorectal cancer.MethodsAll patients with peritoneal metastasis from colorectal cancer who underwent CRS with or without hyperthermic intraperitoneal chemotherapy (HIPEC) between November 2008 and October 2014 were included. The growth inhibition rate was expressed as the ratio between the image density after treatment (T) and that before treatment (control, C). Tumours with a reduction in T/C ratio of less than 20 per cent were defined as resistant and those with a reduction of 20 per cent or more as sensitive. Groups were compared for overall (OS) and disease‐free (DFS) survival.ResultsOf 84 eligible patients, 81 received neoadjuvant chemotherapy (NACT), including 56 patients with an oxaliplatin‐based regimen. Mean(s.d.) follow‐up was 23·4(22·9) months. The median overall survival of all patients was 19·0 (i.q.r. 5·7–36·1) months, with a progression‐free survival time of 10·1 (4·5–17·0) months. Patients who received oxaliplatin‐based NACT had significantly altered chemosensitivity to oxaliplatin; only 20 of 51 such patients showed chemosensitivity to oxaliplatin compared with 16 of 24 who did not undergo oxaliplatin‐based NACT (P = 0·046). However, patients who showed chemoresistance to oxaliplatin had similar OS to those with chemosensitivity (18·8 versus 18·1 months; P = 0·835). The choice of HIPEC agents in patients who received oxaliplatin‐based NACT did not significantly influence survival (oxaliplatin versus mitomycin C: median OS 20·6 (10·9–24·8) versus 19·0 (10·5–34·6) months, P = 0·811; DFS 6·6 (2·8–25·7) versus 9·3 (4·1–13·9) months, P = 0·191).ConclusionPatients who had oxaliplatin‐based NACT showed a higher rate of chemoresistance to oxaliplatin at the time of CRS and HIPEC. The impact of chemosensitivity testing on OS remains unclear and needs further investigation.

The value of GeneXpert MTB/RIF in bronchoalveolar lavage fluid in the diagnosis of smear-negative pulmonary tuberculosis

This study aims to evaluate the diagnostic value of Xpert MTB/ RIF assay in bronchoalveolar lavage fluid (BALF) in subjects with smear-negative pulmonary tuberculosis. From January 2019 to December 2019, 197 patients with suspected pulmonary tuberculosis were recruited, and bronchoalveolar lavage fluid was collected for acid-fast staining smear, liquid culture of Mycobacterium combined drug sensitivity and Xpert MTB/RIF detection. The sensitivity, specificity, positive predictive value and negative predictive value of Xpert MTB/RIF in bronchoalveolar lavage fluid (BALF) were calculated with smear-negative pulmonary tuberculosis as the reference standard. The consistency of xpert MTB/RIF in the diagnosis of rifampicin resistance was evaluated, with the results of Mycobacterium liquid culture drug sensitivity test and drug sensitivity test as the gold standards. The results showed that among 197 suspected tuberculosis patients, 55 patients were not diagnosed with tuberculosis and 142 patients were diagnosed with smear-negative pulmonary tuberculosis. One hundred and twenty three cases (86.62%) were positive for Xpert MTB/ RIF in bronchoalveolar lavage fluid, 15 cases (10.56%) were positive by acid-fast staining smear method, and 88 cases (61.97%) were positive by the liquid culture method. The positive rate of Xpert MTB / RIF was 93.18% (82 / 88), which was higher than that of 75.93% (41 / 54) of the negative BALF mycobacterium culture (χ 2 = 8.598, P&lt;0.01). The sensitivity and specificity of Xpert MTB/RIF for rifampicin resistance were 100.00% and 97.30%, respectively. Therefore, the diagnostic value of Xpert MTB/RIF in bronchoalveolar lavage fluid for bacterialnegative pulmonary tuberculosis is superior to the acid-fast staining smear of lavage fluid and the mycobacterium culture method.

Optimal Contact Concentration of Paclitaxel in the Collagen Gel Droplet-Embedded Culture Drug Sensitivity Test for Human Oral Squamous Cell Carcinoma and Evaluation of Combination with Cetuximab

Objective: A combination of the taxane anticancer drug paclitaxel (PTX) and molecular target drug cetuximab (cMab) is effective for the treatment of head and neck squamous cell carcinoma (HNSCC). However, its use is associated with serious side effects, such as neuropathy and myelosuppression. In addition, it is administered regardless of patient sensitivity because biomarkers indicating its efficacy are unavailable. Therefore, we investigated the usefulness of setting the indicated contact concentration of PTX and predicted the antitumor effect of combined contact with cMab using the collagen gel droplet-embedded culture drug sensitivity test (CD-DST). Method: Twelve human oral squamous cell carcinoma (OSCC) cell lines (i.e., SAS, HSC-2, HSC-3, HSC-4, OSC-19, OSC-20, KON, HO-1-N-1, HO-1-u-1, SAT, SCC-4, and Nialym) were used. Using the CD-DST, we calculated the optimal contact concentration of the cells with PTX based on the clinical response rate of HNSCC and evaluated the combined contact with cMab. Furthermore, nude mice were treated with standalone PTX and PTX + cMab, and the results were compared with those of the CD-DST. Results: Based on the CD-DST, 0.1 μg/mL was the optimal contact concentration of PTX, to which the cells showed dose-dependent sensitivity. Moreover, the CD-DST method was used to evaluate the antitumor effects on OSCC even when PTX was used in combination with cMab. The antitumor effects in the CD-DST and nude mice were correlated (p < 0.05). Conclusion: The CD-DST results suggested that it was possible to predict the clinical effects of single-contact PTX and the enhancing effect of cMab + PTX.

One Step Forward: Successful End-of-Treatment Outcomes of Patients With Drug-Resistant Tuberculosis Who Received Concomitant Bedaquiline and Delamanid in Mumbai, India.

The Médecins Sans Frontières Clinic in Mumbai, India, has been providing concomitant bedaquiline (BDQ) and delamanid (DLM) in treatment regimen for patients with drug-resistant tuberculosis (DR-TB) and limited therapeutic options, referred from other healthcare institutions, since 2016. The study documents the end-of-treatment outcomes, culture-conversion rates, and serious adverse events (SAEs) during treatment. This was a retrospective cohort study based on routinely collected program data. In clinic, treatment regimens are designed based on culture drug sensitivity test patterns and previous drug exposures, and are provided for 20-22 months. BDQ and DLM are extended beyond 24 weeks as off-label use. Patients who initiated DR-TB treatment including BDQ and DLM (concomitantly for at least 4 weeks) during February 2016-February 2018 were included. Of the 70 patients included, the median age was 25 (interquartile range [IQR], 22-32) years and 56% were females. All except 1 were fluoroquinolone resistant. The median duration of exposure to BDQ and DLM was 77 (IQR, 43-96) weeks. Thirty-nine episodes of SAEs were reported among 30 (43%) patients, including 5 instances of QTc prolongation, assessed as possibly related to BDQ and/or DLM. The majority (69%) had culture conversion before 24 weeks of treatment. In 61 (87%), use of BDQ and DLM was extended beyond 24 weeks. Successful end-of-treatment outcomes were reported in 49 (70%) patients. The successful treatment outcomes of this cohort show that regimens including concomitant BDQ and DLM for longer than 24 weeks are effective and can be safely administered on an ambulatory basis. National TB programs globally should scale up access to life-saving DR-TB regimens with new drugs.

Phase Ib study of gemcitabine, nab-paclitaxel, and ficlatuzumab in patients with advanced pancreatic cancer.

693 Background: Paired-related homeodomain transcription factor 1 (Prrx1) isoforms are involved in pancreatic development, pancreatitis, and carcinogenesis. Hepatocyte growth factor (HGF) is a transcriptional target of Prrx1b. Ficlatuzumab is a recombinant humanized HGF antibody, that neutralizes HGF/c-Met binding and HGF-induced c-Met phosphorylation. In preclinical pancreatic ductal adenocarcinoma (PDAC) models, inhibition of Prrx1b-HGF signaling using ficlatuzumab and gemcitabine reduced primary tumor volume and eliminated metastatic disease. Methods: Patients (pts) with previously-untreated metastatic PDAC enrolled in a phase Ib dose escalation study with 3+3 design and two dose cohorts of ficlatuzumab (10mg/kg and 20mg/kg) administered intravenously every other week with gemcitabine (G; 1000mg/m2) and nab-paclitaxel (A; 125mg/m2) given 3 weeks on and 1 week off. This was followed by an expansion phase at the maximally tolerated dose (MTD) of the combination. Results: 24 pts (sex, 12M:12F; median age, 69 years [range, 51-82 years]) were enrolled. No dose-limiting toxicities were identified in the phase 1b (N = 6 pts) and ficlatuzumab at 20mg/kg with GA was advanced to the expansion phase (N = 18 pts). By RECISTv1.1 in the full study population, 7 (29%) pts had partial response, 15 (63%) had stable disease, and 2 (8%) could not be evaluated. Median progression-free survival was 8 months (range, 3-16 months), 4 pts are still on study treatment. The primary toxicities attributed to ficlatuzumab included hypoalbuminemia (grade 3, 21%; any grade, 91%) and edema (grade 3, 8%; any grade, 91%). Nine (38%) of the 24 pts discontinued study treatment due to these toxicities prior to disease progression. Conclusions: The combination of ficlatuzumab with gemcitabine and nab-paclitaxel is associated with durable treatment responses but also significant hypoalbuminemia and edema that may impair treatment tolerability. Serial blood samples were collected for circulating HGF measurements, and mandatory pretreatment biopsies were collected for tumor c-MET pathway activation and 3D organoid culture drug sensitivity testing. Clinical trial information: NCT03316599.

Examination of Factors Influencing the Success Rates of an In Vitro Chemosensitivity Test for Lung Cancer Using Collagen Gel Droplet-Embedded Cultures

Objective: We evaluated the success rates and factors influencing the success rates of the Collagen Gel Droplet-Embedded Culture Drug Sensitivity Test (CD-DST) for pleural effusate in lung cancer patients with carcinomatous pleurisy.Materials and methods: Thirty-two patients with lung cancer having carcinomatous pleurisy were enrolled in this study between January 2014 and May 2017. Their pleural effusions were analysed by CD-DST, and the subjects were classified into two groups: successful cases and unsuccessful cases, based on whether the test was able to successfully determine drug sensitivity. We investigated whether the properties of the pleural effusate might influence the success rate of CD-DST. We observed that the CD-DST tended to have good success rates in patients with higher pH, higher percentage of lymphocytes, higher cell counts, and lower percentage of neutrophils in their pleural effusate. The red blood cell count was higher in the effusate of unsuccessful than successful cases.Conclusion: Our results suggest that the diagnostic success rates of CD-DST for pleural effusion from patients with lung cancer might improve by appropriate sample selection.

Identification of the optimal cetuximab concentration that is effective against oral squamous cell carcinoma in collagen gel droplet embedded culture drug sensitivity testing.

Anticancer drug sensitivity testing using the collagen gel droplet embedded culture drug sensitivity test (CD-DST) on oral squamous cell carcinoma (OSCC) samples beginning from 2010 has been conducted. The present study investigated the effect of adding cetuximab (Erbitux®), a molecularly targeted drug, on anticancer drug activity against clinical OSCC specimens. A total of 25 specimens were obtained from 25 patients with OSCC between October 2013 and December 2017. The present study conducted anticancer drug sensitivity testing for cisplatin (CDDP), 5-fluorouracil (5-FU), cetuximab, three-drug combination, single agent and multi drug combinations, and cetuximab addition to the aforementioned regimens using CD-DST. In addition, the optimum concentration of each drug was evaluated. The overall evaluation success rate of the CD-DST method for OSCC specimens was 84.0% (21 of 25 cases); sensitivity to anticancer drugs and cetuximab could be evaluated. The in vitro efficacy rate of a cetuximab single agent and CDDP + 5-FU (PF) at a cut-off value of 50% was similar to the known clinical response rate. However, at a cut-off value of 50%, the in vitro efficacy of PF + cetuximab was calculated to be 40%, which was higher than the clinical response rate. The CD-DST method could be used to evaluate cetuximab, a molecularly targeted drug. Furthermore, its additive effect on conventional chemotherapy could be evaluated. The CD-DST method is suitable for evaluating and selecting chemotherapy regimens, including molecularly targeted drugs. Future studies are required to generate and evaluate relevant clinical data.

Anterior Gradient 3 Promotes Breast Cancer Development and Chemotherapy Response.

PurposeAnterior gradient 3 (AGR3) belongs to human anterior gradient (AGR) family. The function of AGR3 on cancer remains unknown. This research aimed to investigate if AGR3 had prognostic values in invasive ductal carcinoma (IDC) of breast cancer and could promote tumor progression.Materials and MethodsAGR3 expression was detected in breast benign lesions, ductal carcinoma in situ and IDC by immunohistochemistry analysis. AGR3’s correlations with clinicopathological features and prognosis of IDC patients were analyzed. By cell function experiments, collagen gel droplet-embedded culture drug sensitivity test and cytotoxic analysis, AGR3’s impacts on proliferation, invasion ability, and chemotherapeutic drug sensitivity of breast cancer cells were also detected.ResultsAGR3 was up-regulated in luminal subtype of histological grade I-II of IDC patients and positively correlated with high risks of recurrence and distant metastasis. AGR3 high expression could lead to bone or liver metastasis and predict poor prognosis of luminal B. In cell lines, AGR3 could promote proliferation and invasion ability of breast cancer cells which were consistent with clinical analysis. Besides, AGR3 could indicate poor prognosis of breast cancer patients treated with taxane but a favorable prognosis with 5-fluoropyrimidines. And breast cancer cells with AGR3 high expression were resistant to taxane but sensitive to 5-fluoropyrimidines.ConclusionAGR3 might be a potential prognostic indicator in luminal B subtype of IDC patients of histological grade I-II. And patients with AGR3 high expression should be treated with chemotherapy regimens consisting of 5-fluoropyrimidines but no taxane.

Collagen gel droplet-embedded culture drug sensitivity test (CD-DST) predicts the effect of adjuvant chemotherapy on pancreatic cancer.

We evaluated the clinical effectiveness of collagen gel droplet-embedded culture drug sensitivity tests (CD-DSTs) in predicting the efficacy of adjuvant chemo-therapeutic treatments for pancreatic cancer (PC). The clinicopathological characteristics and prognoses of 22 PC patients who underwent CD-DST after pancreatectomy at Tohoku University between 2012 and 2016 were analyzed retrospectively. Eligibility criteria were resectable or borderline resectable PC, successful evaluation for 5-fluorouracil sensitivity by CD-DST, treatment with S-1 adjuvant chemotherapy, and no preoperative chemotherapy. The rate of successful evaluation by CD-DST was 52.3% in PC. The optimal T/C ratio, defined as the ratio of the number of cancer cells in the treatment group (T) to that in the control group (C), for 5-fluorouracil was 85% using receiver operating characteristic curve analysis. The sensitive group (T/C ratio < 85%; n = 11) had a better recurrence-free survival rate than the resistant group (T/C ratio ≥ 85%; n = 11; P = 0.029). A Cox proportional hazards regression model demonstrated that sensitivity to 5-fluorouracil was an independent predictor of recurrence on multivariate analysis (hazard ratio 3.28; 95.0% CI 1.20-9.84; P = 0.020). CD-DSTs helped to predict PC recurrence after S-1 adjuvant chemotherapy.

A retrospective study on the role of automated real-time nucleic acid amplification technology for rapid and simultaneous detection of mycobacterium tuberculosis and rifampicin resistance in testing different types of tissue specimens of spinal tuberculosis

Objective To compare the role of automated real-time nucleic acid amplification technology for rapid and simultaneous detection of mycobacterium tuberculosis and rifampicin resistance (Xpert MTB/RIF) in testing different types of tissue specimens of spinal tuberculosis. Methods The pus, granulation tissue and caseous necrotic tissue of 223 patients who were diagnosed as having spinal tuberculosis (TB) and received curettage at the Department of Orthopedics of Tianjin Haihe Hospital were collected for bacterial culture and Xpert MTB/RIF assay. Also, the bacterial culture and phenotypic drug-sensitivity test were adopted as the golden standards to calculate the sensitivity and specificity of Xpert MTB/RIF assay. Results The positive rate of Xpert MTB/RIF assays on spinal TB patients’ tissue specimens was higher than that of bacterial culture [ (68.61±7.35)% vs. (44.39±6.51)%, P=0.000]. The positive rate of pus was the highest, followed by granulation tissue, and the caseous necrotic tissue was the lowest by both methods with statistically significant difference [bacterial culture: (44.44±7.08)% > (43.86±7.44)% > (30.72±7.31%), P=0.017; Xpert MTB/RIF assay: (73.02±6.33)% > (63.74±7.21)% > (39.87±7.76%), P=0.000]. The positive rate of Xpert MTB/RIF assay was higher than that of bacterial culture in all three types of specimens (pus and granulation tissue: P=0.000, with statistically significant difference; caseous necrotic tissue: P=0.131, without statistically significant difference). The sensitivity of Xpert MTB/RIF assay on mycobacterium tuberculosis was up to (96.97±3.38)% while the sensitivity (97.44%) and specificity (94.74%) of drug-sensitivity test also remained relatively high. Conclusion For efficient and accurate diagnosis of TB and drug-resistance test and timely provision of effective treatment, multiple specimens, especially the pus of spinal TB patients, should be collected for Xpert MTB/RIF assays. Key words: Spinal tuberculosis; Molecular diagnostics; Mycobacterium tuberculosis

Optimal Cetuximab Contact Concentration Using a Collagen Gel Droplet- Embedded Culture Drug Chemosensitivity Test in Human Oral Squamous Carcinoma Cell Lines

Objective: There is no established therapy for recurrent/metastatic head and neck squamous cell carcinoma (HNSCC), making decisions regarding anticancer agent use and other treatments difficult. Cetuximab, an anticancer drug used as molecular targeted therapy, is administered irrespective of patient sensitivity due to unavailability of biomarkers predicting its efficacy. We investigated the utility of antitumor effect prediction of cetuximab using collagen gel droplet-embedded culture drug sensitivity test (CD-DST).Methods: We evaluated 13 human oral squamous carcinoma cell lines (Ca9-22, SAS, SAT, HSC-2, HSC-3, HSC-4, OSC-19, OSC-20, HO-1-N-1, HO-1-u-1, KON, SCC-4, and Nialym). The expression of cetuximab-related genes was confirmed in each cancer cell line using RT-PCR, and CD-DST was used to measure cell line sensitivity to cetuximab and to calculate an optimal contact concentration based on the HNSCC clinical response rate and efficacy rate. Furthermore, CD-DST was performed at this calculated optimal contact concentration using the treatment regimens cetuximab+cisplatin and cetuximab+cisplatin+5-fluorouracil. In vivo assessment was performed in nude mice that were administered cetuximab (250 μg/ml) and cisplatin alone and in combination, and the results were compared with the CD-DST results. Results: Results of RT-PCR showed no specificity. With CD-DST, the optimal contact concentration of cetuximab was 250 µg/ml. An enhanced antitumor effect was observed in some of the cell lines with low susceptibility to CDDP and CF. Antitumor effects for CD-DST and nude mouse experiments were almost equal.Conclusions: CD-DST can indicate the clinical efficacy of cetuximab. Regarding anticancer drug groups, low-sensitivity cell lines exhibited high sensitivity for regimen including cetuximab, suggesting cetuximab may enhance the antitumor effects of existing cytotoxic anticancer drugs.

Clinical study on collagen gel droplet-embedded culture drug sensitivity test for multidrug combination chemotherapy and super selective intra-arterial infusion chemoradiotherapy in oral squamous cell carcinoma.

Using trace three-dimensional culture, the collagen gel droplet-embedded culture drug sensitivity test (CD-DST) can be tested even in cases with a small number of cells, including oral squamous cell carcinoma (OSCC), and evaluation of the antitumor effect with a drug concentration close to the in vivo level is possible. The present report aimed to evaluate the utility of the CD-DST in the assessment of the in vitro efficacy of single-agent and multidrug combination chemotherapy for OSCC in comparison with the clinical response rates and to examine the possible clinical application of CD-DST for such cases. A total of 33 OSCC patients from whom 33 samples were obtained from January 2010 to September 2015 were included. CD-DST was performed, individually and in combination, on the three drugs [i.e., cisplatin (CDDP), 5-fluorouracil (5-FU), and docetaxel (DOC)] and on super selective intra-arterial infusion chemoradiotherapy (IACRT). The overall evaluable rate of the CD-DST in OSCC was 81.8% (27 of 33 cases) and the sensitivity to each anticancer drug was evaluated. The in vitro efficacy rates of IACRT, cisplatin + 5-fluorouracil, and docetaxel + cisplatin + 5-fluorouracil (TPF) confirmed the estimated clinical response rates. In 14 of 33 patients, the results of CD-DST were compared with clinical efficacy, which was judged based on measurable lesions on imaging. For TPF therapy, the sensitivity test of the IACRT had a positive predictive value of 90.9% (10 of 11 cases) and a negative predictive value of 100% (3 of 3 cases); the accuracy of the susceptibility test for the anticancer agents was 92.8% (13 of 14 cases). The CD-DST may be useful in selecting multidrug combination chemotherapy and IACRT for OSCC, however, accumulation of further clinical data is required in the future.

Impact of the individualization of the first-line chemotherapy for advanced colorectal cancer based on collagen gel droplet-embedded drug sensitivity test.

Leucovorin (FOL) and fluorouracil (5-FU) plus oxaliplatin (l-OHP; FOLFOX) or FOL and 5-FU plus irinotecan (SN-38; FOLFIRI) are widely used as first-line chemotherapy regimens in the treatment of advanced colorectal cancer (CRC). However, second-line chemotherapy must be abandoned in certain cases due to disease progression, adverse effects or high medical cost. Therefore, the most effective regimen should be selected as first-line chemotherapy. We reported that individualization of first-line treatment (FOLFOX/FOLFIRI/Dual/Poor responder) was possible using the collagen gel droplet-embedded culture drug sensitivity test (CD-DST) and that individualized first-line chemotherapy with CD-DST may improve the prognosis of patients with unresectable CRC. The aim of the present prospective cohort study was to evaluate the individualization of first-line chemotherapy using CD-DST, with a focus on prognosis. Between March 2008 and December 2015, tumor specimens were obtained from 120 patients with CRC who had not received preoperative chemotherapy. CD-DST was performed and the growth inhibition rate (IR) was determined by exposure for 24 h with 5-FU and l-OHP (6.0 and 3.0 µg/ml, respectively) and 5-FU and SN-38 (6.0 and 0.2 µg/ml, respectively). The cumulative distribution of IR values under each condition was evaluated on the basis that the clinical response to FOLFOX and FOLFIRI is equivalent (~50%). The prognosis of dual responder was improved compared with that of poor responders, however this difference was identified to be significant. There was no different prognosis between patients treated with an appropriate first-line regimen and patients treated with an inappropriate first-line regimen in dual responders. However, in poor responders, there were significant differences of prognosis between patients treated with an appropriate first-line regimen and patients treated with an inappropriate first-line regimen (P=0.036). In conclusion, the results from the present study suggest that administration of the recommended first-line regimen using CD-DST for patients with unresectable CRC is important for the improvement of prognosis, particularly in poor responders.