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"Difteri adalah penyakit infeksi akut saluran nafas atas yang disebabkan oleh Corynebacterium diphtheria, bersifat sangat menular dengan tingkat kematian tinggi. Beberapa tahun terakhir ditemukan kembali kasus difteri di berbagai wilayah Indonesia, walaupun program vaksinasi telah dilaksanakan secara luas. Gejala difteri yang ditimbulkan sebagian besar disebabkan oleh toksin dan dapat berakibat fatal. Kerusakan yang disebabkan oleh toksin yang telah terikat pada sel host tidak dapat diperbaiki walaupun telah diberikan antitoksin. Beberapa laporan menunjukkan bahwa pemberian antitoksin pada hari pertama pengobatan dapat mempengaruhi angka kematian kurang dari 1%, dan dapat meningkat sampai 30% bila ditunda sampai hari ke 6. Medium diagnosis cepat (MDC) yang akan dikembangkan dalam penelitian ini akan menggabungkan tiga pemeriksaan mikrobiologi dalam satu langkah, yaitu inokulasi, uji biokimia dan serologi. Penelitian ini merupakan eksperimen laboratorium, yang menggunakan strain referensi dan isolat tersimpan dari Balitbangkes Kemkes RI. Penelitian ini meliputi tiga tahap yaitu : optimasi konsentrasi larutan telurit, komposisi medium, dan konsentrasi Fosfomisin. Hasil pertumbuhan pada MDC memperlihatkan gambaran halo pada strain yang potensial toksigenik dan presipitat pada strain toksigenik. Waktu pemeriksaan pada MDC sekitar 24-48 jam lebih cepat dari pada prosedur standar.

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Diphtheria is an acute infectious disease of upper respiratory tract caused by Corynebacterium diphtheria, and highly contagious with high mortality rate. Few years back cases of diphtheriae were found in various parts of Indonesia, although vaccination programs have been widely implemented. Symptoms of diphtheria largely caused by toxins and can be fatal. Damage caused by toxin that has been bound to the host cell can not be repaired dispite of administration of antitoksin.

Some reports showed that administration of antitoksin on the first day of treatment affected the mortality rate, i.e. less than 1% compared to 30% when delayed until day 6. Medium Diagnosis Cepat (MDC) established in this research will integrate three microbiology examination in one step i.e. inoculation, biochemical and serology tests.

This investigation was a laboratory experiment, using reference strains and isolates stored in The National Research and Development of The Ministry of Health Republik of Indonesia. This study covered three stages is follow : optimization of tellurite solution concentration, medium composition, Fosfomycin concentration.

The results showed halo formation of potentially toxigenic strains and precipitate of toxigenic strain on MDC. The examination MDC was faster than the standard procedure in took about 24-48 hours."

"Latar belakang: Difteri merupakan penyakit infeksi endemis dan menjadi masalah kesehatan di Indonesia. Pada tahun 2017 kejadian luar biasa difteri terjadi di beberapa provinsi di Indonesia. Tujuan: Untuk mengetahui prediktor kematian difteri klinis pada anak di Provinsi DKI Jakarta dan Kabupaten Tangerang selama kejadian luar biasa tahun 2017-2018. Metode: Penelitian kohort retrospektif dilakukan di lima rumah sakit rujukan di DKI Jakarta dan satu di Kabupaten Tangerang periode 1 Januari 2017-31 Agustus 2018. Pasien anak usia 1-18 tahun dengan diagnosis difteri klinis dinilai faktor prediktor yang berhubungan dengan luaran kematian. Uji korelasi chi square dilakukan untuk mengetahui hubungan variabel bebas dan luaran. Multivariat analisis dilakukan untuk menentukan prediktor kematian. Analisis data dilakukan dengan program SPSS for Window ver 20,0. Hasil: Pasien anak dengan difteri klinis sejumlah 283 kasus dengan case fatality rate 3,5%. Riwayat imunisasi dasar (RR 6,967; p 0,003), suara serak (RR 7,611; p 0,035), stridor (RR 16,963; p<0,001), bullneck (RR 28,456; p<0,001), limfadenopati (RR 3,838; p 0,045), komplikasi miokarditis (p<0,001), leukositosis >15.000sel/mm3 (RR 7,500; p 0,004), trombositopenia (RR 35,549; p<0,001), kultur C. diphtheriae positif (RR 6,587; p 0,04) berhubungan dengan kematian. Analisis multivariat menunjukkan stridor (HR 11,951; p 0,006), lekositosis (HR 11,425; p 0,01), dan trombositopenia (HR 44,199; p<0,001) berhubungan dengan kematian. Simpulan: Stridor, lekositosis dan trmbositopenia merupakan faktor prediktor kematian pada difteri klinis anak.

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Background: Diphtheria is endemic in Indonesia. In 2017 diphtheria outbreak has taken place in several provinces.

Objective: To identify predictors of mortality of pediatric patients with clinical diphtheria during 2017-2018 outbreak in the Province of Jakarta and Tangerang District.

Methods: A retrospective cohort study has been held at five referral hospitals in the Province of Jakarta and one in Tangerang District during January 2017 and 31 August 2018. The study group is children age group of 1-18 years old admitted with sign and symptoms and discharge as clinical diphtheria. All details that is demographic data, clinical features, immunization status, complication and laboratory profiles and outcome were analysed. Variables were compared among survivors and non survivors to determine the predictors of mortality. A chi square test and cox regression was done to assess association between variables and outcome. Data were analysed using SPSS for Window ver 20,0.

Results: A total of 283 pediatric patients with clinical diphtheria were included in the study group with case fatality rate of 3.5%. Basic immunization status (RR 6.967; p0.003), hoarseness (RR 7.611; p0.035), stridor (RR 16.963; p<0.001), bullneck (RR 28.456; p<0.001), limphadenopaty (RR 3.838; p0.045), myocarditis (p<0,001), leukocytosis >15,000 cell/mm3 (RR 7.500; p0.004), thrombocytopenia <150,000 cell/mm3 (RR 35.549; p<0.001), C. diphtheriae positive culture (RR 6.587; p0.04) were found correlated to mortality. Multivariat analysis showed that stridor (HR 11.951; p0.006), leukocytosis (HR 11,425; p0,01), and thrombocytopenia (HR 44.199; p<0.001) correlated to death.

Conclusion: Diphtheria is fatal disease with increased mortality. Presence of stridor, leukocytosis and thrmbocytopenia are important predictors of mortality."

"Dari tahun 2005 sampai dengan tahun 2006 telah terjadi Kejadian Luar Biasa (KLB) difteri di Kabupaten Tasikmalaya pada kelompok umur 1 ? 15 tahun sebanyak 55 anak (15 kasus meninggal, AR = 0,45% dan CFR = 31,91%). Pada Januari 2007 juga telah terjadi KLB difteri di Kabupaten Garut pada kelompok umur kasus 2 ? 14 tahun sebanyak 17 anak (2 kasus meningal, CFR = 11,76%, AR = 1,5%). Penelitian ini bertujuan untuk mengetahui hubungan lingkungan rumah dengan kejadian difteri pada Kejadian Luar Biasa (KLB) difteri tersebut. Penelitian menggunakan desain kasus kontrol. Kasus berasal dari 15 desa lokasi KLB difteri sebanyak 72 anak dan kontrol berasal dari 1 desa terpilih secara random yang bukan dari kecamatan lokasi KLB difteri sebanyak 72 anak. Pengumpulan data dilakukan dengan cara wawancara dengan ibu anak pada kelompok kasus maupun kelompok kontrol menggunakan kuesioner untuk mendapatkan data lingkungan rumah, sumber penularan, status imunisasi dan pengetahuan ibu. Hasil analisis multivariat menunjukkan bahwa variabel yang berhubungan bermakna dengan kejadian difteri adalah kepadatan hunian ruang tidur, kelembaban dalam rumah, jenis lantai rumah, sumber penularan, status imunisasi dan pengetahuan ibu. Disimpulkan bahwa lingkungan rumah, pengetahuan ibu dan sumber penularan bukanlah faktor utama yang mempengaruhi terjadinya difteri, sedangkan yang paling dominan dalam mempengaruhi kejadian difteri adalah status imunisasi, yaitu risiko terjadinya difteri pada anak dengan status imunisasi DPT/DT yang tidak lengkap 46,403 kali lebih besar dibandingkan dengan anak dengan status imunisasi yang lengkap. Untuk itu cakupan program imunisasi hendaknya makin ditingkatkan sehingga semua anak terlindungi oleh imunisasi difteri.Since 2005 up to 2006 diphtheria out break had occur in Tasimalaya District among 1 ? 15 year old children. Total cases are 55 children with cases died with the Case Fatality Rate (CFR) 31.91%. Further on, January 2007 the same out break occur in Garut District, with 17 cases and 2 cases died (CFR 11.76%). Research objective is to identify the correlation of housing environmental condition with the diphtheria out break. Design study was case control study. The amount of 72 cases had taken from the 15 villages on the out break areas and the same amount (72) non cases taken from the village out of the out break areas. Data were collected through interviewed with structure questioner with the mother as the respondent. Data collected were housing environment, the source of infection, immunization status, and mother knowledge concerning the diphtheria. Research conclude that factors involved in diphtheria out break are housing member room density, housing humidity, quality of the floor, the source of the infection, immunization status of the children, and mother knowledge about the disease. The importance factors for the diphtheria out break are immunization status, with the OR of 46.403 greater of non immunization children compare with those had immunization. Therefore immunization program should be further intensified in order to give fully diphtheria protection for the hole children population in those areas."

"ABSTRACTLow adult vaccination coverage in Indonesia may contribute to a recent outbreak of diphtheria in Indonesia. Although well known as a pediatric vaccine, diphtheria vaccination should be administered as booster to adolescence and adults for longer prevention. Adult vaccine differs from pediatric vaccine but have similar protection. Additionally, there is special recommendation to vaccinate pregnant women and elderly people aged 65 years or more."

"Latar Belakang: Patogenitas Corynebacterium pada infeksi difteri, sangat terkait dengan toksin yang dihasilkannya. Indonesia merupakan negara ke dua di dunia yang memiliki kasus difteri terbanyak dengan 1.665 kasus dan 29 kasus kematian di tahun 2018. Toksin difteri dapat dihasilkan oleh 3 spesies penyebab, yaitu C. diphtheriae, C. ulcerans, dan C. pseudotuberculosis yang sulit dibedakan secara mikroskopik dan kultur, sehingga diperlukan deteksi molekuler untuk membedakanya. Karakterisasi C. diphtheriae sebagai spesies penyebab utama, diperlukan untuk mengetahui hubungan kekerabatanya dengan spesies negara lain untuk mencari kemungkinan sumber infeksi. Metode: Sebanyak 108 sampel klinis digunakan dalam penenlitian ini. Teknik kultur dilakukan untuk mendeteksi 3 Corynebacterium patogenik, sedangkan realtime PCR hanya dirancang untuk mendeteksi C. ulcerans dan C. pseudotuberculosis. Pengujian inhibitor, sensitifitas, dan spesifisitas dilakukan pada tahap optimasi. Karakterisasi C. diphtheriae dilakukan dengan metode sekuensing menggunakan gen rpoB parsial pada kultur sampel klinis. Hasil: Limit deteksi real-time PCR untuk C. ulcerans dan C. pseudotuberculosis secara berurutan sebanyak 4,49 dan 1,06 DNA copy number. Uji spesifisitas terhadap 18 mikrorganisme menunjukkan tidak terdapat reaksi silang. Pengujian terhadap 108 sampel klinis memberikan hasil yang sama dengan kultur, tidak ditemukan C. ulcerans dan C. pseudotuberculosis. Pada kultur sampel klinis ditemukan C. diphtheriae sebanyak 10 sampel (9,26%), yang dapat dikelompokkan menjadi 4 clade yaitu clade I, III, IV dan V dengan similaritas 99,2 % sampai 99,7%. Kesimpulan: Kasus suspek difteri dalam studi ini tidak berkaitan dengan infeksi C. ulcerans dan C. pseudotuberculosis, dan hanya positif C. diphtheriae . Hasil karakterisasi gen rpoB pada C. diphtheriae, memperlihatkan hubungan kekerabatan dengan beberapa negara.

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Background: Pathogenicity of Corynebacterium in diphtheria infection is closely related to toxin production. Indonesia is the second highest country in the world that has the most diphtheria cases with 1,665 cases and 29 deaths in 2018. Diphtheria toxin can be produced by 3 species, such as C. diphtheriae, C. ulcerans and C. pseudotuberculosis which are difficult to distinguish microscopically and culture, therefor molecular detection is needed to differentiate them. Characterization of C. diphtheriae as the main causative species, is needed to determine its relationship with other countries to find possible source of infection.

Method: A total of 108 clinical samples were used in this study. Culture techniques were performed to detect 3 pathogenic Corynebacterium and real-time PCR was only designed to detect C. ulcerans and C. pseudotuberculosis. Inhibitor, sensitivity and specificity testing were carried out at the optimization stage. Characterization of C. diphtheriae from culture of clinical samples, was carried out by sequencing method using partial rpoB genes.

Result: The real-time PCR detection limits for C. ulcerans and C. pseudotuberculosis were 4.49 and 1.06 DNA copy number, respectively. Specificity test for 18 microorganism showed no cross reaction. Tested on 108 clinical samples gave the same results as culture, there were not found C. ulcerans and C. pseudotuberculosis. In clinical culture samples found 10 (9.26%) C. diphtheriae, which can be grouped into 4 clades namely clades I, III, IV and V with similarities of 99.2% to 99.7%.

Conclusion: Suspected diphtheria cases in this study were not related to C. ulcerans and C. pseudotuberculosis infections, and were only positive for C. diphtheriae. The results of the rpoB gene characterization test on C. diphtheriae showed a close relationship with several countries.

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"Difteri merupakan penyakit re-emerging yang disebabkan oleh bakteri Corynebacterium diphtheria dan menyebabkan kematian. Kematian akibat difteri ini di dunia memiliki CFR 5 – 10% dan di Indonesia CFR difteri sebesar 2% sehingga masih menjadi masalah kesehatan masyarakat. Penelitian ini bertujuan untuk mengetahui faktor-faktor yang berhubungan dengan kematian akibat difteri di Indonesia Tahun 2018. Penelitian ini menggunakan data surveilans laporan rutin kasus difteri di Indonesia tahun 2018. Desain studi yang digunakan adalah cross-sectional dengan unit analisis individu yang menderita difteri di Indonesia tahun 2018. Pada penelitian ini didapatkan 817 kasus dengan 2,8% penderita meninggal dan tempat tinggal responden paling banyak di Jawa Barat, yaitu 211 penderita. Faktor jenis kelamin, status imunisasi primer difteri, dan status pemberian Anti Difteri Serum (ADS) tidak berhubungan secara statistik dengan kematian akibat difteri. Faktor yang berhubungan secara statistik dengan kematian akibat difteri, yaitu umur < 15 tahun (OR = 7,863; 95% CI = 1,831 – 33,77) dan diagnosis penderita difteri konfirmasi laboratorium (OR = 2,774; 95% CI = 1,000 – 7,693).

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Diphtheria is a re-emerging disease caused by the Corynebacterium diphtheria bacteria and causes death. The death from diphtheria in the world has CFR of 5-10% while in Indonesia has CFR is 2% that make it becomes one of public health concerns. This study aims to determine the factors associated with diphtheria deaths in Indonesia in 2018 using surveillance data on routine reports of diphtheria cases in Indonesia in 2018. The study design used was cross-sectional with a unit of analysis of individuals suffering from diphtheria in Indonesia in 2018. In this study there were 817 cases with 2.8% of patients dying and mostly of them living in West Java province (211 patients). Sex factors, primary diphtheria immunization status, and Serum Anti Diphtheria (ADS) status were not statistically related to diphtheria deaths. Factors statistically associated with diphtheria deaths were age <15 years (OR = 7.863; 95% CI = 1,831 - 33,77) and diagnosis of diphtheria patients from laboratory confirmation (OR = 2,774; 95% CI = 1,000 - 7,693)."

"Penyakit difteri sudah jarang di temukan di Indonesia. Sebagaimana terlihat dari hasil Survei Kesehatan Rumah Tangga (SKRT) tahun 1986 dan 1992 sebagai penyebab kesakitan dan kematian pada bayi/balita menduduki peringkat 5 (9,4 %) dan 6 (3,3 %) di Jawa - Bali serta tahun 1995 peringkat ke 7 (2,1 %), namun dalam kurun waktu Oktober tahun 2000 sampai dengan Juli tahun 2001 penyakit ini menimbulkan kejadian luar biasa (KLB) pada beberapa Desa dan Kecamatan di Kabupaten Cianjur Jawa Barat.Dari data-data yang ada bahwa program imunisasi DPT di tempat KLB terjadi, berjalan dengan lancar bahkan Desa tersebut sudah mencapai status Universal Child Immunization (UCI). Sejalan dengan kontradiksi tersebut penulis terdorong untuk melakukan penelitian terhadap faktor-faktor risiko yang berhubungan dengan kejadian difteri selain faktor imunisasi.Dari beberapa faktor tersebut penulis memfokuskan pada faktor kepadatan serumah karena cara penularan penyakit ini adalah dengan kontak langsung, kontak tidak langsung melalui benda, tangan, makanan dan minuman yang terkontaminasi. Disamping cara penularannya faktor-faktor lain tersebut juga berkorelasi positif terhadap hubungan kepadatan serumah dengan kejadian difteri. Jumlah sampel dalam penelitian ini adalah 186 orang dengan jumlah kasus 93 orang dan kontrol 93 orang.Hasil dari penelitian ini didapat bahwa faktor kepadatan serumah berhubungan dengan kejadian difteri dan faktor yang berpengaruh terhadap hubungan tersebut adalah faktor kepadatan human kamar tidur, sumber pengeluaran, faktor pengeluaran keluarga, faktor status imunisasi dan faktor status gizi.Berdasarkan hasil penelitian disarankan kepada Dinas Kesehatan Kabupaten Cianjur untuk tetap mempertahankan cakupan imunisasi yang tinggi secara terus menerus khususnya DPTIDT dan merata di semua Desa khususnya yang padat penghuninya. Alga disarankan kepada Puskesmas ditempat yang penduduknya padat untuk mengaktifkan pemberian makanan tambahan melalui Posyandu, upaya perbaikan gizi keluarga di Kecamatan dan melalui program pemberian makanan tambahan di sekolah. Kepada Pemerintah Kabupaten Cianjur disarankan untuk mengalokasikan dana Program Pengembangan Kecamatan (PPK) untuk pembangunan perumahan khususnya penduduk yang padat penghuninya.Daftar bacaan : 45 (1964 - 2041)

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Relation between Crowded in House and Difteri on Outbreak Difteri in Cianjur District West Java year 2000 - 2001Difteri has been rarely found in Indonesia. As shown on the Survey result of household health (SKRT) in year 1986 and 1992, the disease are causes the death for baby and pre school, and having its ranks of 5`h (9,4 %) and 6`s (3,3 %) in Java - Bali also in the year of 1995 is the 7' ranks (2,1 %) out of Java - Bali. While in October 2000 until July 2001 there is created an out break affect in several Village and Sub District in Cianjur District West Java.

From the existing datas, program of DPTlDT Immunization in out break place had happened is good and that Villages have get Universal Child Immunization (UCI) status. According to this contradictif the writer motivated to take the research about risk factor to relation with difteri occurred that difteri beside the immunization factors. From several factors, here with the Writer focus on crowded in house due to this disease is contamination of direct contact, contamination of indirect contact trough the body, arms, food and drinks.

Besides the contamination, other factors are also correlation positive towards the crowded in house with difteri occurred. Total samples in these examination are 186 poeples with 93 person case group and 93 person control groups.

Result of this research founded that crowded in house factors related with the difteri disease and another risk factor affected to that relation is : crowded in bedroom, contamination facts, family expenses, immunization status and nutrition status.

Based on the result of this research, the writer suggested to the Govennent in Cianjur expected to alocated compensation load of oil and gas priority to do the Program Pengembangan Kecamatan (PPK) in the matter of houses construction especially in the crowded area. To Department of Health in Cianjur District to keep continiuosly the immunization of DPTIDT in all Villages which having a high population. Also suggested to Public Health Centre to increase service give meal supplement in Integreted Health Post (Posyandu) and by Upaya Perbaikan Gizi Keluarga (UPGK) programme at Sub District and by Pemberian Makanan Tambahan di Sekolah (PMTS) programme. It is also recommended to another researcher to investigate the relation crowded in house and difteri occurred by applying cohort design, it is expected that all information found in the research may help the stakeholders to implement the policies related to the recovery of difteri disease.

Bibliography 45 (1964 - 2001)"

"ABSTRACTBackground: in recent years, diphtheria has reemerged in several countries including Venezuela, Yemen, Bangladesh, and Haiti. Similarly, Indonesia also showed an increased number of diphtheria cases in 2010-2017 despite the Diphteria, Tetanus, Pertussis (DTP) immunization program applied in Indonesia for children. This study aimed to evaluate the epidemiology of diphtheria cases which occurred in Indonesia during 2010-2017. Methods: this was a retrospective study of diphtheria cases in Indonesia. The following source of data about diphtheria disease burden and vaccine coverage was obtained from Ministry of Health Republic of Indonesia, Indonesian Pediatric Society and World Health Organization South East Asia Regional Office. Results: the number of diphtheria cases in Indonesia were distributed across 30 provinces with a total of 811 cases in 2011; 1,192 cases in 2012; 296 cases in 2014; and 939 cases in 2017. Based on age group, the highest number of case fatality rate were in age group of 5-9 years old. Diphtheria immunization coverage in Indonesia among children was fluctuated, reported as 67.7 % in 2007, 61.9 % in 2010, 75.6% in 2013 and 61.3% in 2018. In addition to that, the organization of internal medicine has recommend booster of DPT immunization every 10 years for those children that had received complete DPT vaccination during childhood, however this was not applied. As the countermeasure towards this trend, the Ministry of Health implemented three rounds of Outbreak Response Immunization (ORI) targeted for the age group of 0-1-6 months old and 1-18 years old in 2017 and tailor approached for adults that had exposed to cases. Banten, DKI Jakarta and West Java were the first three provinces to implement this program considering their condensed population and high risk of disease transmission. Conclusion: in Indonesia, there was dramatic increase of diphtheria case in 2010-2017, where immunization in children should be reinforced by increasing coverage more than 95% and adult boosted vaccination approaches should be initiated to prevent the spread of these fatal diphtheria diseases in Indonesia."

"Latar belakang: Infeksi C. diphtheriae toksigenik memerlukan penanganan segera karena dapat mengancam nyawa. Pemeriksaan kultur sebagai uji baku emas hanya dapat dilakukan di laboratorium rujukan dan memakan waktu lama. Pemeriksaan awal yang dapat dilakukan adalah menggunakan teknik pewarnaan. Pewarnaan khusus difteri seperti Albert atau Neisser dapat menggambarkan granul metakromatik yang spesifik terhadap Corynebacterium sp., namun laboratorium yang mampu mengerjakannya sangat terbatas. Oleh karena itu, berbagai laboratorium menggunakan teknik pewarnaan Gram sebagai alternatif untuk menunjang diagnosis dini difteri.Tujuan: Evaluasi pemeriksaan mikroskopik langsung yang rutin dilakukan pada spesimen swab tenggorok untuk menunjang diagnosis dini difteri.Metode: Uji diagnostik dengan desain potong lintang. Pemeriksaan mikroskopik Gram dan pewarnaan khusus Albert serta Neisser dibandingkan dengan kultur sebagai baku emas untuk mendapatkan nilai sensitivitas, spesifisitas, nilai duga positif, nilai duga negatif, dan akurasi. Uji kesesuaian antara kedua teknik pewarnaan dan PCR dinilai menggunakan koefisien Kappa. Analisis multivariat dilakukan pada variabel independen data faktor risiko dan gambaran klinis. Nilai probabilitas yang dihasilkan dimasukkan ke dalam kurva Receiver Operating Characteristic (ROC). Pertambahan luas Area Under Curve (AUC) dinilai setelah menambahkan pemeriksaan mikroskopik ke dalam model.Hasil: Pewarnaan khusus difteri dan Gram memiliki sensitivitas 70% dan 20%, spesifisitas 83,72% dan 79,06%, nilai duga positif 33,33% dan 10%, nilai duga negatif 96% dan 89,47% serta akurasi 82,29% dan 72,92%. Kesesuaian minimal antara kedua metode pewarnaan (reliabilitas 4-15%). Pewarnaan khusus difteri memiliki kesesuaian yang lebih baik dengan uji PCR,dibandingkan dengan pewarnaan Gram. Pewarnaan khusus difteri menunjukan penambahan nilai diagnostik sebesar 3,8% pada model faktor risiko dan gambaran klinis yang bermakna untuk menunjang diagnosis dini difteri.Kesimpulan: Pewarnaan khusus difteri pada spesimen klinis langsung untuk diagnosis dini difteri tidak dapat digantikan dengan pewarnaan Gram. .....Background: Infection caused by toxigenic C. diphtheriae requires immediate treatment because it can be life threatening. Culture examination as gold standard can only be done in a reference laboratory and is time-consuming. The initial examination can be performed using the staining technique. Diphtheria special staining such as Albert or Neisser can characterize metachromatic granules that are specific to Corynebacterium sp., but the availability of these stains in many laboratories are limited. Therefore most laboratories use Gram staining technique as an alternative to support early diagnosis of diphtheria.Objective: Evaluation of the direct microscopic examination commonly used on throat swab specimens to support the early diagnosis of diphtheria.Methods: Diagnostic test with a cross-sectional design. Gram microscopic examination and special staining of Albert and Neisser were compared with culture examination as the gold standard to obtain sensitivity, specificity, positive predictive value, negative predictive value, and accuracy. The realibility test between the two staining and PCR techniques was assessed using the Kappa coefficient. Multivariate analysis was performed on the independent variables of risk factors and clinical features data. The resulting probability value was entered into the Receiver Operating Characteristic (ROC) curve. The improvement of the area under the curve (AUC) was assessed after microscopic examination was added on the model.Results: Diphtheria and Gram staining had a sensitivity of 70% and 20%, specificity 83.72% and 79.06%, positive predictive value 33.33% and 10%, negative predictive value 96% and 89.47% and accuracy 82.29 % and 72.92%. Minimal concordance was found between the two staining methods (4-15%). Diphtheria-specific staining have better realibility result, compared to the PCR test, than Gram staining. Diphtheria special staining showed an additional 3.8% diagnostic value, in the significant of risk factors and clinical features model, to support the early diagnosis of diphtheria.Conclusion: Diphtheria special staining directly on clinical specimens for early diagnosis of diphtheria cannot be replaced by Gram staining."

"Beberapa jenis penyakit membutuhkan penatalaksanaan secara cepat dan tepat guna menurunkan risiko fatal penderita, salah satu diantaranya adalah difteri. Waktu sangat berharga untuk bisa menolong penderita karena keterlambatan penatalaksanaan bisa meningkatkan kematian kasus hingga 20 kali lipat. Di sisi lain, metode diagnostik konvensional sebagai gold standard membutuhkan waktu 3?5 hari sehingga diperlukan metode diagnostik alternatif untuk membantu menegakkan diagnosis difteri. Direct polymerase chain reaction (PCR) dapat menjawab tantangan atas besarnya biaya dan lamanya waktu yang dibutuhkan untuk pemeriksaan laboratorium. Penelitian bertujuan untuk mengembangkan metode direct PCR sebagi metode alternatif diagnostik difteri secara cepat, mudah dan hemat. Sebanyak 15 sampel yang terdiri dari 10 isolat Corynebacterium diphtheriae toksigenik dan 3 Corynebacterium non-diphtheriae nontoksigenik ditambah dengan 2 spesimen klinis (usap tenggorok) digunakan untuk optimasi metode direct PCR dibandingkan dengan PCR standard sebagai kontrol. Hasil penelitian menunjukkan bahwa direct PCR dapat digunakan untuk deteksi dan amplifikasi gen target dengan benar seperti halnya PCR standard sehingga pada seluruh sampel C. diphtheriae tampak pita pada 168 bp (penanda gen dtxR) dan 551 bp (penanda gen tox). Sebaliknya pada sampel lain tidak tampak pita pada kedua tempat tersebut. Direct PCR dapat mendeteksi sel bakteri sampai dengan 71 CFU/uL. Oleh karena itu, disimpulkan bahwa direct PCR dapat digunakan sebagai metode diagnostik alternatif untuk membantu menegakkan diagnosis difteri secara cepat, mudah dan hemat.

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Direct PCR: Alternative Diagnostic Method for Diagnosis of Diphtheria Rapidly, Easily and Cost Effective. Some diseases require immediate and appropriate treatment to decrease the fatality risk patients incident, for example diphtheria. Time to help patients is very crucial since delay of therapy may increase the mortality cases up to 20 times. In other hands, conventional diagnostic methods (the gold standard) for diagnosis of diphtheria is time consuming and laborious. Therefore, an alternative diagnostic method which is rapid, easy and inexpensive is needed. In this case, direct PCR has been proved to reduce time and cost in laboratory examination. This study aimed to develop direct PCR as alternative diagnostic method for diagnosis of diphtheria rapidly, easily, and inexpensive. Fifteen samples include 10 isolates of Corynebacterium diphtheriae (toxigenic) and 3 isolates of Corynebacterium non- diphtheriae (nontoxigenic) and 2 clinical specimens (throat swab) was examined by performing direct PCR method and a standard PCR method was used for optimizing the protocols. Result showed that direct PCR can be used to amplify target genes correctly as well as standard PCR. All of C. diphtheriae samples showed bands at 168 bp (dtxR gene marker) and 551 bp (tox gene marker) while no band appeared in others. Direct PCR detected at least 71 CFU/uL of bacterial cells in samples. We concluded that direct PCR can be used for alternative diagnostic method for diagnosis of diphtheria which is rapid, easy and cost effective."